Antimicrobial Compositions and Methods of Use Thereof

ABSTRACT

The invention provides antimicrobial compositions comprising one or more acid and one or more organic diol. In one embodiment, the invention&#39;s compositions have an acidic pH. The compositions may optionally further contain one or more oxidizing agent (including stabilized oxidizing agent and/or unstabilized oxidizing agent), and/or one or more surfactant. In particular embodiments, the acid lacks one or both of —NH group and-NH 2  group. The invention&#39;s compositions are useful in all stages of handling agricultural products, in hospitals, and in commercial and household applications.

This application claims priority under 35 U.S.C. §119(e) to U.S.Provisional Application Ser. No. 61/024,558 to Kel Eugene Lemons, filedJan. 30, 2008, herein incorporated by reference in its entirety for allpurposes.

FIELD OF THE INVENTION

The invention provides antimicrobial compositions comprising one or moreacid and one or more organic diol. In one embodiment, the invention'scompositions have an acidic pH. The compositions may optionally furthercontain one or more oxidizing agent (including stabilized oxidizingagent and/or unstabilized oxidizing agent), and/or one or moresurfactant. In particular embodiments, the acid lacks one or both of —NHgroup and —NH₂ group. The invention's compositions are useful in allstages of preparation of agricultural products, in hospitals, and incommercial and household applications.

BACKGROUND OF THE INVENTION

Microbial contamination of food, in hospitals, in animals, as well as incommercial and residential buildings continues to be a problem. Inparticular, pathogenic microbial contamination of fruits and vegetableshas been on the rise, including contamination of fruits and vegetablesin the field, during harvesting, transport and/or processing, bypathogenic microbes from animals, humans, organic fertilizer and/orwater supply. Bacteria are estimated to cause some 24 million cases ofdiarrheal disease annually in the U.S. Over 40,000 cases ofcontamination by Salmonella were reported from 1983 to 1987 inlaboratory surveillance data.

Several compositions and methods are available for reducing microbialcontamination (e.g., Bailey et al., U.S. Pat. No. T964,007 and U.S. Pat.No. 4,074,058; Faergemann et al., WO/89/12469; Andrews et al., U.S. Pat.No. 5,490,992; Estrada, U.S. Pat. No. 5,599,571; Andrews, U.S. Pat. No.5,569,461; Bautista et al., U.S. Pat. No. 6,045,846; Conners et al.,U.S. Pat. No. 6,086,833; Bender et al., U.S. Pat. No. 6,287,617; Koefodet al., U.S. Pat. No. 7,090,882; Howarth et al., U.S. Pat. No.7,182,966; Shane et al., U.S. Patent Application US 2002/0134317; andHilgren et al., U.S. Patent Application US 2007/0098751), includingthose that contain oxidizers. However, prior art compositions thatcontain oxidizers continue to show low antimicrobial activity in thepresence of organic contaminants, and especially at the lowertemperatures used for processing, storing and transporting agriculturalproducts. Also, the prior art's compositions have the additionalproblems of increased foaming and of the oxidizer gassing out of thesolution due to bio-load and/or too low or too high pH, thus adverselyimpacting active ingredients and worker safety. Furthermore, higherlevels of oxidizers are required to reduce bio-load and turbidity of thesolutions used for antimicrobial treatment, which raises safety concernswhen ingesting products that are treated with those compositions. Inaddition, the presence of plant material in the prior art solutionsreduces the longevity of the oxidizer, thus reducing its antimicrobialactivity and increasing the cost of production because of the need toreplenish the oxidizer often.

Thus, there remains a need for improved compositions and methods forreducing microbial contamination.

SUMMARY OF THE INVENTION

The invention provides an aqueous composition comprising an (a) acidthat does not contain an —NH group and an —NH₂ group, and (b) organicdiol, wherein the composition (i) has acidic pH and (ii) hasantimicrobial activity. In a particular embodiment, the amount of theacid alone and the organic diol alone has lower antimicrobial activitycompared to antimicrobial activity of a combination of the acid and theorganic diol.

While not intending to limit the type of acid in any of the invention'scompositions, in one embodiment, the acid comprises an organic acid, asexemplified by, but not limited to, citric acid, ascorbic acid, lacticacid, malic acid, octenic acid, oxalic acid, ursolic acid,hydroxyethanoic acid and salts of each of the organic acid. In anotherembodiment, the acid comprises an inorganic acid as exemplified by, butnot limited to, ortho-phosphoric acid, chromic acid, hydrobromic acid,hydrochloric acid, nitric acid, sulfuric acid, and salts of each of theinorganic acid. In yet a further embodiment, the acid is exemplified by,but not limited to, acetic acid, adipic acid, benzoic acid, glutaricacid, isoascorbic acid, lactic acid, mandelic acid, phosphoric acid,propionic acid, salicylic acid, sorbic acid, succinic acid, tartaricacid, sodium acid pyrophosphate, acidic sodium hexametaphosphate,ethylenediaminetetraacetic acid and salts of each of the acid. Withoutintending to limit the amount of the acid in any of the invention'scompositions, in one embodiment, the composition comprises from 0.001wgt. % to 40 wgt. % of the acid. In one embodiment, at least one acid inthe composition is generally recognized as safe (GRAS).

It is not intended that the type and/or amount of organic diol in any ofthe invention's compositions be limited to a particular type and/oramount. Nonetheless, in one embodiment, the organic diol comprises analiphatic diol, such as, without limitation, a simple diol. In oneembodiment, the organic diol is exemplified by, but not limited to,propylene glycol, ethylene glycol, 1,3-butanediol, 1,7-heptanediol,1,2-octanediol, 1,5-pentanediol and polyethylene glycol. In oneembodiment, the composition comprises from 0.001 wgt. % to 5 wgt. % ofthe organic diol. In one embodiment, at least one organic diol in thecomposition is generally recognized as safe (GRAS).

In some embodiments, the composition further comprises an oxidizingagent, such as, but not limited to, an unstabilized oxidizing agent.While not intending to limit the oxidizing agent (whether stabilized orunstabilized) to a particular type and/or amount, in one embodiment, theoxidizing agent is exemplified by, but not limited to, bromine, sodiumhyporchlorite, calcium hyporchlorite, chlorine dioxide, chlorine,hypochlorous acid, hydrogen peroxide, peroxyacetic acid, and ozone. Inanother embodiment, the composition comprises from 0.001 wgt. % to 30wgt. % of the oxidizing agent.

In particular embodiments, the composition further comprises asurfactant. While not intending to restrict the type of surfactant, inone embodiment, the surfactant comprises an anionic surfactantexemplified by, but not limited to, sodium lauryl sulfate, ammoniumlauryl sulfate, alkylbenzene sulfonic acid, sodium 2-ethylhexyl sulfate,and dioctyl sodium sulfosuccinate. In an alternative embodiment, thesurfactant comprises a neutral surfactant exemplified by, but notlimited to, octyl phenol ethoxylate, glyceryl monostearate,polyglyceryl-10 decaoleate, and lauryl lactyl lactate. In yet a furtherembodiment, the surfactant comprises a cationic surfactant exemplifiedby, but not limited to, cetrimonium bromide, cetylpyridinium chloride,benzalkonium chloride, cocamidopropyl betaine, ethylene oxide moiety,and propylene oxide moiety. Still in other embodiments, the surfactantcomprises a linear alkylbenzene sulfonate, alcohol sulfate, alpha-olefinsulfonate, alcohol ethoxylate, nonylphenyl ethoxylate,alkylpolyglucoside, fatty alkanoamide, fatty amine oxide, sodiumdioctylsulfosuccinate, dodecylbenzene sulfonic acid, dodecylbenzenesulfonic acid salt, sulfonated oleic acid sodium salt, sodiumdodecylbenzene sulfonate, dodecyidiphenyloxidedisulfonic acid, anddodecyidiphenyloxidedisulfonic acid salt. It is not contemplated thatthe invention be limited to a particular concentration of surfactant.However, in some embodiments, the composition comprises from 0.001 wgt.% to 0.1 wgt. % of the surfactant.

Also without intending to limit the invention's compositions to anyparticular combination of acid, organic diol, oxidizing agent, andsurfactant, in one embodiment, the composition is exemplified by, butnot limited to, compositions AA, BB, CC, DD, xx1, xx2, xx3, and xx4. Theinvention also provides a concentrated solution and a diluted solutionof any of the compositions disclosed herein, such as of compositions AA,BB, CC, DD, xx1, xx2, xx3, and xx4. In a particular embodiment,

It is not intended to limit the temperature of the composition. Thus, insome embodiments, the composition is at a temperature from the freezingtemperature of the composition to 120° C.

The invention additionally provides an aqueous composition comprising ana) acid, b) organic diol, and c) unstabilized oxidizing agent, whereinthe composition (i) has acidic pH, and (ii) has antimicrobial activity.In a particular embodiment, the antimicrobial activity of (a) the acid,(b) the organic diol, (c) the oxidizing, (d) a combination of the acidand of the organic diol, (e) a combination of the acid and of theoxidizing agent, and (f) a combination of the organic diol and of theoxidizing agent, is lower than antimicrobial activity of a combinationof the acid, of the organic diol and of the oxidizing agent. Inparticular embodiments, the composition further comprises d) asurfactant.

While not intending to limit the invention's compositions to anyparticular combination of acid, organic diol, unstabilized oxidizingagent and surfactant, in some embodiments, the composition isexemplified by, but not limited to, compositions A, B, C, D, E, F, G, H,I, J, K, L, M, N, O, x1a, x1, x2, x3, x4, x5, x6, x7, x8, x9, x10, x11,x12, x13, x14, x15, a1, a2, a3, a4 and a5. The invention additionallycontemplates a concentrated solution and a diluted solution of any ofthe compositions disclosed herein. In some embodiments, at least one ofthe acid, the organic diol and the oxidizing agent is generallyrecognized as safe (GRAS).

In certain embodiments, (a) the oxidizing agent comprises hypochlorousacid, and (b) the rate of decrease in concentration of the hypochlorousacid in the presence of the diol is greater than in the absence of thediol. Preferred, but not limiting, embodiments, include those in whichantimicrobial activity in the presence of the diol is greater than inthe absence of the diol. In certain embodiments, the acidic pH is frompH 2 to pH 5.5. In yet other embodiments, the composition is at atemperature from the freezing temperature of the composition to 120° C.

The invention also provides an agricultural product having a surfacefilm that comprises any of the compositions disclosed herein, includingan a) acid that does not contain an —NH group and an —NH₂ group, and b)organic diol. In some embodiments, the film further comprises anoxidizing agent, such as an unstabilized oxidizing agent. In otherembodiments, the film further comprises a surfactant. Certainembodiments include those where the film is edible, dry appearing and/orclear. In an alternative embodiment, the agricultural product ispre-harvest or post-harvest.

The invention additionally provides an agricultural product having asurface film that comprises an a) acid, b) organic diol, and c)unstabilized oxidizing agent. In some embodiments, the film furthercomprises d) a surfactant.

Also provided by the invention is a method for making an aqueousantimicrobial composition comprising an (a) acid that does not containan —NH group and an —NH₂ group, and (b) organic diol, wherein thecomposition (i) has acidic pH and (ii) has antimicrobial activity. Inone embodiment, the method comprises a) providing i) an acid that doesnot contain an —NH group and an —NH₂ group, and ii) an organic diol, andb) mixing the acid and the diol in water to produce an aqueousantimicrobial composition. In one embodiment, the method furthercomprises c) mixing an oxidizing agent in the aqueous antimicrobialcomposition. In particular embodiments, mixing with the oxidizing agentis in the absence or presence of an oxidizing agent stabilizer. In analternative embodiment, the method further comprises c) mixing asurfactant in the aqueous antimicrobial composition. The invention alsoprovides an aqueous antimicrobial composition produced by any of themethods described herein.

The invention additionally provides a method for making an aqueousantimicrobial composition comprising an a) acid, b) organic diol, and c)unstabilized oxidizing agent, wherein the composition (i) has acidic pH,and (ii) has antimicrobial activity. In one embodiments, the methodcomprises a) providing an i) acid, ii) organic diol, and iii)unstabilized oxidizing agent, b) mixing the acid and the diol in waterto produce a first mixture, and c) mixing the oxidizing agent with thefirst mixture to produce an aqueous antimicrobial composition. Inparticular embodiments, the method further comprises mixing a surfactantin the aqueous antimicrobial composition. In alternative embodiments,the oxidizing agent comprises chlorine. The invention also provides anaqueous antimicrobial composition produced by any of the methodsdisclosed herein.

In yet another embodiment, the invention provides a method for reducingthe number of microbes on a surface, comprising a) providing i) a firstsurface comprising a first number of microbes, and ii) anantimicrobially effective amount of any of the compositions describedherein, and b) contacting the first surface with the composition underconditions that produce a contacted surface comprising a reduced numberof the microbes compared to the first number of the microbes on thefirst surface in the absence of the contacting. In a particularembodiment, the composition comprises an (a) acid that does not containan —NH group and an —NH₂ group, and (b) organic diol, wherein thecomposition (i) has acidic pH and (ii) has antimicrobial activity. Thecomposition may further comprise an oxidizing agent, such as anunstabilized oxidizing agent, and/or a surfactant.

While not intending to limit the antimicrobial activity of any of theinvention's compositions, in one embodiment, (A) the first surfacecomprises from 3-log to 8.78-log number of the microbes, (B) thecontacting is for at least 2 minutes at a temperature of at least 34°F., and (C) the reduced number of the microbes comprises at least a3-log reduction in the number of the microbes on the contacted surfacecompared to the first surface. In an alternative embodiment, the reducednumber of the microbes comprises 100% reduction in the number of themicrobes on the contacted surface compared to the first number of themicrobes on the first surface.

It is not intended that the invention be limited to a particularantimicrobial effect on pathogenic versus non-pathogenic microbes.Nonetheless, in one embodiment, the first surface comprises a pathogenicmicrobe and a non-pathogenic microbe, and the reduced number of themicrobes comprises a greater reduction in the number of the pathogenicmicrobe than in the number of the non-pathogenic microbe. In anotherembodiment, the pathogenic microbe and the non-pathogenic microbecomprise Gram-positive bacteria. In a particular embodiment, thepathogenic Gram-positive bacteria comprise Staphylococcus aureus and thenon-pathogenic Gram-positive bacteria comprise Lactobacillus sp. In yetanother alternative embodiment, the pathogenic microbe comprisesGram-negative bacteria and the non-pathogenic microbe comprisesGram-positive bacteria. Alternatively, the pathogenic Gram-negativebacteria are exemplified by, but not limited to, Escherichia coli andSalmonella enterica, and the non-pathogenic Gram-positive bacteriacomprise Lactobacillus sp. In a further alternative, the pathogenicmicrobe comprises pathogenic bacteria and the non-pathogenic microbecomprises non-pathogenic bacteriophage. In yet another embodiment, thepathogenic bacteria comprise Escherichia coli and the non-pathogenicbacteriophage comprises bacteriophage T4. In a further embodiment, thepathogenic microbe comprises pathogenic virus and the non-pathogenicmicrobe comprises non-pathogenic bacteriophage. In another alternativeembodiment, the pathogenic virus comprises Rhinovirus and thenon-pathogenic bacteriophage comprises bacteriophage T4.

The invention's methods are not limited to the nature or number of stepsother than the step of contacting a surface with the invention'scompositions. Nonetheless, the invention's methods that use acomposition comprising an (a) acid that does not contain an —NH groupand an —NH₂ group, and (b) organic diol, wherein the composition (i) hasacidic pH and (ii) has antimicrobial activity, may further comprise stepc) drying the contacted surface to produce a dried surface comprisingone or more of the acid and the organic diol. In an alternativeembodiment, the method comprises c) washing the contacted surface underconditions that reduce the amount of one or more of the acid and theorganic diol on the contacted surface.

Alternatively, the invention's methods that use a composition comprisingan a) acid, b) organic diol, and c) unstabilized oxidizing agent,wherein the composition (i) has acidic pH, and (ii) has antimicrobialactivity, may further comprise step c) drying the contacted surface toproduce a dried surface comprising one or more of the acid, the organicdiol, and the oxidizing agent. In an alternative embodiment, the methodfurther comprises c) washing the contacted surface under conditions thatreduce the amount of one or more of the acid, the organic diol, and theoxidizing agent on the contacted surface.

It is contemplated that any of the invention's methods wherein thesurface is a surface of an agricultural product, the method furthercomprises c) storing the contacted agricultural product. In someembodiments, storing is at a chilling temperature, such as from 30° F.to 45° F.

The temperature at which the surface is contacted with the invention'scompositions is not limited to any particular temperature. Thus, in oneembodiment, contacting is at a temperature from the freezing temperatureof the composition to 120° C.

Without limiting the type of surface that is contacted with theinvention's compositions, in a particular embodiment, the surface is asurface of an agricultural product and the contacted surface has reduceddiscoloration compared to the first surface in the absence of thecontacting. In a particular embodiment, the discoloration comprisespinking and the agricultural product is exemplified by, but not limitedto, lettuce, cabbage, celery, and Bok Choy. In an alternativeembodiment, the discoloration comprises browning and the agriculturalproduct is exemplified by, but not limited to, lettuce, cabbage, celery,Bok Choy, potato, parsnip, avocado, apple, strawberry, spinach, meat,poultry and seafood.

The invention does not contemplate limiting the method of contacting asurface with the invention's compositions. However, in one embodiment,contacting comprises one or more of immersing, dipping, spraying,fogging, aerosoling, washing, and scrubbing the surface with thecomposition. In a particular embodiment, the contacting does not alterone or more of odor, texture and color of the surface.

It is not intended that the invention be limited to the type or sourceof the microbe against which the invention's compositions haveantimicrobial activity. Thus, in one embodiment, the microbe comprisesbacteria. In a particular embodiment, the bacteria compriseGram-negative bacteria exemplified by, but not limited to, Escherichiacoli, Salmonella, Pseudomonas, Moraxella, Helicobacter,Stenotrophomonas, Bdellovibrio, acetic acid bacteria, Legionella,alpha-proteobacteria, Hemophilus influenzae, Klebsiella pneumoniae,Legionella pneumophila, Pseudomonas aeruginosa, Proteus mirabilis,Enterobacter cloacae, Serratia marcescens, Helicobacter pylori,Salmonella enteritidis, Salmonella typhi, Campylobacter, andAcinetobacter baumanii. In another embodiment, the bacteria compriseGram-positive bacteria exemplified by, but not limited to, Bacillus,Listeria, Staphylococcus, Streptococcus, Enterococcus, Clostridium, andMollicutes. In particularly preferred embodiments, the Gram-positivebacteria are selected from the group of Listeria monocytogenes,Staphylococcus aureus and Lactobacillus sp.

In an alternative embodiment, the microbe comprises a fungus. In aparticular embodiment, the fungus comprises a flagellated fungusselected from the group of Plasmodiophoromycetes, Chytridiomycota andOomycetes. In an alternative embodiment, the fungus comprises anon-flagellated fungus selected from the group of Zygomycota,Ascomycota, Deuteromycetes and Basidiomycetes. In yet a furtherembodiment, fungus is selected from the group of Aspergillus flavus,Septoria lycopersici, and Stachybotrys chartarum.

In yet another alternative embodiment, the microbe comprises a virus. Ina particular embodiment, the virus is selected from the group ofrhinovirus, human papilloma virus, human immunodeficiency virus,hepatitis virus, Newcastle disease virus, cardiovirus, corticoviridae,cystoviridae, epstein-barr virus, filoviridae, hepadnviridae, hepatitisvirus, herpes virus, influenza virus, inoviridae, iridoviridae,metapneumovirus, orthomyxoviridae, papovavirus, paramyxoviridae,parvoviridae, polydnaviridae, poxyviridae, reoviridae, rhabdoviridae,semliki forest virus, tetraviridae, toroviridae, vaccinia virus, andvesicular stomatitis virus. In another embodiment, the virus comprises abacteriophage selected from the group of Myoviridae, Siphoviridae,Podoviridae, Tectiviridae, Corticoviridae, Lipothrixviridae,Plasmaviridae, Rudiviridae, Fuselloviridae, Inoviridae, Microviridae,Leviviridae, and Cystoviridae. In particular embodiments, thebacteriophage comprises bacteriophage T4.

In a further embodiment, the microbe comprises a nematode, asexemplified by Aphelenchoides fragariae.

The invention is not limited to the type or source of surface that maybe contacted with the invention's compositions. Thus in one embodiment,the surface is a surface of an agricultural product, exemplified by, butnot limited to, vegetable, fruit, bulb, root, berry, herb, seed, meat,poultry, seafood, poultry egg, animal hide, and feather. In anotherembodiment, the surface is a surface of skin and/or of an inanimateobject.

The invention further contemplates an agricultural product comprising acontacted surface that is produced by the steps of any of the methodsdescribed herein.

Also provided by the invention is a method for reducing discoloration ofan agricultural product, comprising a) providing i) an agriculturalproduct having a first surface, ii) an anti-discoloration effectiveamount of any of the compositions disclosed herein, and b) contactingthe agricultural product with the composition under conditions thatproduce a contacted surface having reduced discoloration compared todiscoloration of the first surface in the absence of the contacting.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the level of hypochlorous acid in solution, as determinedby the DPD assay. (A) Composition “x1a” containing ortho-phosphoric acid0.03 wgt. %, propylene glycol 0.02 wgt. %, hypochlorous acid 0.005 wgt.% at pH 4. (B) Chlorine solution containing ortho-phosphoric acid 0.001wgt. % and hypochlorous acid 0.005 wgt. % at pH 7.

DEFINITIONS

To facilitate understanding of the invention, a number of terms aredefined below.

The term “wgt. %” refers to the relative weight of two compounds whenexpressed as a percentage. For example, a solution containing 10 wgt. %of ortho-phosphoric acid propylene glycol dissolved in water refers to asolution prepared by diluting 10 grams of ortho-phosphoric acidpropyleneglycol with water to a final weight of 100 grams.

“Logarithm” and “log” of a number, such as when used in reference to aconcentration, cell number, etc., interchangeably refer to the power orexponent to which a base of 10 must be raised in order to produce thenumber. For example, 6-log means 1,000,000. Thus, a 3-log reductionmeans a 1,000 fold reduction.

“Aqueous” solution or compoistion refers to a solution or composition inwhich the solvent is water.

An “organic” chemical compound (e.g., organic acid, organic diol,organic oxidizer, organic solvent, etc.) is a chemical compound thatcontains carbon. In contrast, an “inorganic” chemical compound (e.g.,inorganic acid, inorganic oxidizer, inorganic solvent, etc.) is achemical compound that does not contain carbon.

“Microbe” and “microorganism” interchangeably refer to an organism thatis too small to be seen by the naked human eye. Microbes includeprokaryotes (such as bacteria, and virus (including bacteriophage),etc.), and eukaryotes (such as protists (including nematodes), animals,fungi, and plants). Microbes may be pathogenic or non-pathogenic.

“Pathogen” refers to an organism that causes a disease in amulticellular animal. In contrast, a “non-pathogen” is an organism thatdoes not cause disease in an animal. “Animal” includes mammals (e.g.,humans, non-human primates, murines, ovines, bovines, ruminants,lagomorphs, porcines, caprines, equines, canines, felines, ayes, etc.),avians (e.g., chicken), amphibians (e.g., Xenopus), reptiles, etc.

The term “bacteria” refers to all prokaryotic organisms, including thosewithin all of the phyla in the Kingdom Procaryotae. Bacteria includeMycoplasma, Chlamydia, Actinomyces, Streptomyces, and Rickettsia. Formsof bacteria include cocci, bacilli, spirochetes, spheroplasts,protoplasts, etc. Bacteria may be Gram-negative or Gram-positive.“Gram-negative” and “Gram-positive” refer to staining patterns with theGram-staining process that is well known in the art (Finegold andMartin, Diagnostic Microbiology, 6th Ed. (1982), CV Mosby St. Louis, pp13-15). “Gram-positive bacteria” are bacteria that retain the primarydye used in the Gram-stain, causing the stained cells to appear darkblue to purple under the microscope. “Gram-negative bacteria” do notretain the primary dye used in the Gram-stain, but are stained by thecounterstain. Thus, Gram-negative bacteria appear red.

“Gram-negative bacteria” include the proteobacteria, exemplified byEscherichia coli, Salmonella, and other Enterobacteriaceae, Pseudomonas,Moraxella, Helicobacter, Stenotrophomonas, Bdellovibrio, acetic acidbacteria, Legionella, alpha-proteobacteria such as Wolbachia and manyothers. Gram-negative bacteria include bacilli that cause respiratoryproblems (such as Hemophilus influenzae, Klebsiella pneumoniae,Legionella pneumophila, Pseudomonas aeruginosa), urinary problems(Escherichia coli, Proteus mirabilis, Enterobacter cloacae, Serratiamarcescens), and gastrointestinal problems (Helicobacter pylori,Salmonella enteritidis, Salmonella typhi and Campylobacter).Gram-negative bacteria associated with nosocomial infections includeAcinetobacter baumanii, which cause bacteremia, secondary meningitis,and ventilator-associated pneumonia in intensive care units of hospitalestablishments.

“Gram-positive bacteria” include the phylum Firmicutes (exemplified bythe genera Bacillus, Listeria, Staphylococcus, Streptococcus,Enterococcus, and Clostridium), and the Mollicutes (exemplified byMycoplasma that lack cell walls and cannot be Gram stained). Listeriamonocytogenes is a Gram-positive bacteria that is of particular concernin many ready-to-eat meat, poultry, seafood, and dairy processing chillbrine applications.

The term “virus” refers to obligate, ultramicroscopic, intracellularparasites incapable of autonomous replication (i.e., replicationrequires the use of the host cell's machinery). Viruses are exemplifiedby, but not limited to, rhinovirus, human papilloma virus, humanimmunodeficiency virus, hepatitis virus, Newcastle disease virus,cardiovirus, corticoviridae, cystoviridae, epstein-barr virus,filoviridae, hepadnviridae, herpes virus, influenza virus, inoviridae,iridoviridae, metapneumovirus, orthomyxoviridae, papovavirus,paramyxoviridae, parvoviridae, polydnaviridae, poxyviridae, reoviridae,rhabdoviridae, semliki forest virus, tetraviridae, toroviridae, vacciniavirus, and vesicular stomatitis virus.

A “fungus” is a eukaryotic organism that is a member of the kingdomFungi. Fungi are heterotrophic organisms possessing a chitinous cellwall. The majority of species grow as multicellular filaments calledhyphae forming a mycelium. Some fungal species also grow as singlecells. Fungi include “mold,” which are microscopic fungi that grow inthe form of multicellular filaments, called hyphae. Molds includedermatophytes (i.e., pathogenic fungus that infects the skin)exemplified by the genera Epidermophyton, Microsporum and Trichophyton(such as Trichophyton rubrum and Trichophyton mentagrophytes).Trichophyton can cause tinea pedis (athlete's foot) and/or tinea cruris(jock itch). Plant fungi include flagellated fungi and non-flagellatedfungi.

Flagellated fungi include Plasmodiophoromycetes, Chytridiomycota andOomycetes. Plasmodiophoromycetes include Plasmodiophora brassicae thatcauses club root of cabbage, and Spongospora subterranean that causespowdery scab of potatoes. Chytridiomycota include Olpidium spp. thatinfect pollen, Physoderma maydis that causes brown spot of corn leaves,and Synchytrium spp. that cause wart of potatoes. The Peronosporalesgroup is exemplified by the late blight of potato fungus Phytophthorainfestans, Peronospora, Bremia, Plasmopara and others that cause “downymildews”, the “damping off” fungi, Pythium spp., and the white rustfungi, Albugo spp.

Non-flagellated Fungi include Zygomycota, Ascomycota, Deuteromycetes andBasidiomycetes. Zygomycota include the Mucorales that are exemplified bymembers of the bread mold genus Rhizopus, and Hoanephora that causesblossom blight and decay of squash. Ascomycota include Taphrinadeformans that causes peach leaf curl, and Nematospora that causes seeddecay and root rot on cotton, Plectomycetes such as Ophiostoma(Ceratocystis), and O. ulmi that causes Dutch elm disease, Pyrenomycetessuch as powdery mildews of the genus Erysiphe that is common on grasses,Phyllactinia on oaks and other trees, and Uncinula on grapes and othershrubs, Discomycetes such as Sclerotinia that causes stromatic rot ofvegetables, and Monilinia, the cause of brown rot of peaches, andLoculoascomycetes such as Myriangiales that include Elsinoe species thatcause citrus scab, and Dothideales that include Capnodium species thatcause sooty molds of plants. Deuteromycetes include Acomycetes such asspecies of Alternaria, Bipolaris, Botrytis, Cercospora, Diplodia,Dreschlera, Exerohilum, Fusarium, Phoma, Phomopsis, Rhizoctonia, andVerticillium that cause molds, blights, cankers, leaf spots, and rootrots. Basidiomycetes include Uredinales, Exobasidiales, andAphyllophorales that cause rusts, smuts, felt fungi, root rots, heartrots, and thread-blights.

A “bacteriophage” is a virus that infects bacteria. Bacteriophages areclassified in the families Myoviridae, Siphoviridae, Podoviridae,Tectiviridae, Corticoviridae, Lipothrixviridae, Plasmaviridae,Rudiviridae, Fuselloviridae, Inoviridae, Microviridae, Leviviridae, andCystoviridae

“Nematode” and “roundworm” interchangeably refer to a multicellularorganism that is a member of the Phylum Nematoda. Exemplary nematodesthat are agricultural pests include corn pests such as Belonolaimus(Sting Nematode), Criconemoides (Ring nematode), Helicotylenchus (SpiralNematode), Heterodera zeae (Corn Cyst Nematode), Hoplolaimus (LanceNematode), Xiphinema (Dagger Nematode), Longidorus (Needle Nematode),Meloidogyne (Root-Knot Nematode), Pratylenchus (Lesion Nematode),Paratrichodorus (Stubby-Root Nematode), Tylenchorhynchus (StuntNematode); potato pests such as Meloidogyne chitwoodi (ColumbiaRoot-knot Nematode), Globodera rostochiensis (Golden Nematode),Meloidogyne hapla (Northern Root Knot Nematode), Ditylenchus destructor(Potato Rot Nematode), Globodera pallida (Pale Potato Cyst Nematode);soybean pests such as Heterodera glycines (Soybean cyst nematode) andBelonolaimus spp. (Sting nematode); sugar beet pests such as Heteroderaschachtii (Sugar beet cyst nematode) and Nacobbus aberrans (Falseroot-knot nematode); turf pests such as Belonolaimus species (StingNematode), Hoplolaimus galeatus (Lance Nematode), Meloidogyne species(Root-knot Nematodes) and Criconemoides species (Ring Nematode); treesand vines pests such as Bursaphelenchus xylophilus (Pine wilt nematode),Tylenchulus semipenetrans (Citrus nematode), Radopholus similis(Burrowing nematode), Belonolaimus longicaudatus (Sting nematode),Xiphinema americanum (Dagger nematode), Mesocriconema xenoplax (Ringnematode), Meloidogyne hapla (Root-knot nematode), Tylenchorhynchus spp.(Stunt nematode), Rotylenchulus spp. (Reniform nematode) andPratylenchus spp. (Lesion nematode); ornamentals and garden vegetablespests such as Aphelenchoides spp. (Foliar nematodes), Meloidogyne spp.(Root-knot nematodes), Ditylenchus dipsaci (Stem and bulb nematode), andBelonolaimus longicaudatus (Sting nematode).

The term “surface” refers to one or more of the faces of athree-dimensional object, including surfaces that are visible to thenaked eye (e.g., agricultural products, inanimate objects, etc.) andsurfaces not visible to the naked eye (e.g., open stomata and damagedcells on a plant leaf surface).

“Agricultural product” refers to plant material and/or animal materialthat may be useful to man, as well as products derived directly orindirectly therefrom (e.g., cut flowers, cooked meat, etc.).Agricultural products include food products and non-food products.

“Food product” refers to an agricultural product that is “edible” (i.e.,suitable for use as food) to at least one animal (e.g., human,livestock, etc.). For example, the invention's compositions may beedible when they are present as a film on a food product and when thetype and/or amount of the components are generally recognized as safe(GRAS). Food products include “pre-harvest” and “post-harvest” products.Food products are exemplified by, but not limited to, fruits,vegetables, herbs, seeds, nuts, meat, poultry, seafood, poultry eggs,etc. Food products may be fresh or processed. “Fresh food product”refers to a food product that has not been cooked or frozen (i.e.,exposed to temperatures higher or lower than room temperature, i.e.,ambient temperature). For example, a “fresh” food product includes, butis not limited to a, raw food product, unprocessed food product, foodproduct that has not been heated above ambient temperature, food productthat has not been cooled below ambient temperature, food product thatdoes not contain a preservative, and/or irradiated food product. A“fresh” food product may be “fresh frozen,” “frozen fresh,” and “freshlyfrozen,” which are terms that interchangeably refer to a food productthat is quickly frozen while still fresh. A “fresh” food product may beblanched, i.e., briefly scalded, before freezing to prevent nutrientbreakdown. “Produce” is a fresh food product derived from a plant (e.g.,fruit, vegetable, herb, seed, nut, leaf, stem, bulb). “Processed foodproduct” refers to a food product that has been manipulated by manincluding a product that has been cut, chopped, sliced, peeled, ground,milled, irradiated, frozen, cooked (e, g, blanched and/or pasteurized),homogenized, germinated, washed, colored, waxed, hydro-cooled,refrigerated, shelled, and/or had leaves, stems, or husks removed.

“Non-food product” refers to an agricultural product that is not edibleto at least one animal. Non-food products include cut flowers,houseplants, animal hides, feathers, skin, shells, etc.

“Inanimate object” refers to an object not endowed with life or spiritsuch as equipment that is used for processing and/or harvestingagricultural products, hospital instruments and surfaces, surgical toolsand apparatus, household items, buildings, tunnels, underpasses,overpasses, floors, doors (frames, door knobs, etc.), entrances, exits,ceilings, bathrooms and fixtures, furniture, wood, Teflon™, ultra highmolecular weight (UHMW) polyethylene, ceramic tile, steel (e.g.,stainless steel), cement, glass, paint (e.g., acrylic paint), andreverse osmosis (RO) membrane (i.e., semi permeable thin film compositemembranes, referred to as TFC or TFM).

“Skin” refers to the integument of an animal separated from the body,with or without hair. The animal may be living or non-living. Skin maybe on the outside of the body, tissue on the inside of the body, and/ortissue at or near openings on the body, including openings madesurgically.

“Film” refers to a thin covering, coating or layer. A film may be“clear” i.e., transparent and/or translucent. Alternatively, the filmmay be “opaque.” Films may be visible to the human naked eye orinvisible to the human naked eye. A film may be “edible,” i.e., is nottoxic when ingested by an animal. To illustrate, an edible filmcontaining one or more of the invention's compositions includes, withoutlimitation, a film in which the components are listed as GRAS and/orFCC. A film may be “adherent,” i.e., is not removed from a surface afterrinsing with water for 1 minute. A liquid film may appear dry to thenaked human eye.

“Bio-film” is a film containing an aggregation of microorganisms.Bio-films are characterized by structural heterogeneity, geneticdiversity, complex community interactions, and an extracellular matrixof polymeric substances.

“Bio-load” and “biological load” refer to the waste output of organisms,plant matter, soil, extracts, and any other biological matter that iscapable of changing the oxygenation rate of a fluid, e.g., water.

The terms “reduce,” “decrease,” and grammatical equivalents when inreference to a numerical value (e.g., number of cells, concentration,etc.) of any molecule (e.g., acid, diol, oxidizing agent, surfactant,etc.), and/or phenomenon (e.g., antimicrobial activity, microbialcontamination, cell division, cell viability, discoloration, pinking,browning, etc.) in a first sample relative to a second sample, mean thatthe quantity of the numerical value in the first sample is lower than inthe second sample by any amount that is statistically significant usingany art-accepted statistical method of analysis. The reduction may bedetermined objectively and/or subjectively. In one embodiment, thequantity of the numerical value in the first sample is at least 10%lower than, at least 25% lower than, at least 50% lower than, at least75% lower than, at least 90% lower than, at least 99% lower than, and/or100% lower than the quantity of the numerical value in the secondsample.

The terms “increase,” and grammatical equivalents when in reference to anumerical value (e.g., number of cells, concentration, etc.) of anymolecule (e.g., acid, diol, oxidizing agent, surfactant, etc.), and/orphenomenon (e.g., antimicrobial activity, microbial contamination, celldivision, cell viability, discoloration, pinking, browning, etc.) in afirst sample relative to a second sample, mean that the quantity of thenumerical value in the first sample is higher than in the second sampleby any amount that is statistically significant using any art-acceptedstatistical method of analysis. The increase may be determinedobjectively and/or subjectively. In one embodiment, the quantity of thenumerical value in the first sample is at least 10% greater than, atleast 25% greater than, at least 50% greater than, at least 75% greaterthan,and/or at least 90% greater than, and/or at least 100% greater thanthe quantity of the same molecule and/or phenomenon in the secondsample.

“Acidic pH” refers to a pH less than 7.0, as exemplified by, but notlimited to, a pH less than pH 6.5, less than pH 6.0, less than pH 5.5,less than pH 5.0, less than pH 4.5, less than pH 4.0, less than pH 3.5,less than pH 3.0, less than pH 2.5, less than pH 2.0, and less than pH1.5. Thus, an acidic pH includes a pH from 2.0 to 6.0, from 2.0 to 5.0,from 2.0 to 4.0, and from 2.0 to 3.0.

Unless otherwise indicated, all numbers expressing quantities ofingredients, temperatures, microbes, reaction conditions, and so forthas used herein, are to be understood as being modified in all instancesby the term “about,” which refers to variation in the numerical quantitythat can occur, for example, through typical measuring and liquidhandling procedures used for making concentrates or using solutions inthe real world; through inadvertent error in these procedures; throughdifferences in the manufacture, source, or purity of the ingredientsemployed to make the compositions or carry out the methods; and thelike. Accordingly, unless indicated to the contrary, the numericalparameters herein are approximations that may vary depending upon thedesired properties sought to be obtained by the present invention.Notwithstanding that the numerical ranges and parameters describing thebroad scope of the invention are approximations, the numerical values inthe specific examples are reported as precisely as possible. Anynumerical value, however, inherently contains standard deviations thatnecessarily result from the errors found in the numerical value'stesting measurements.

DESCRIPTION OF THE INVENTION

The invention provides antimicrobial compositions comprising one or moreacid and one or more organic diol. In one embodiment, the invention'scompositions have an acidic pH. The compositions may optionally furthercontain one or more oxidizing agent (including stabilized oxidizingagent and/or unstabilized oxidizing agent), and/or one or moresurfactant. In particular embodiments, the acid lacks one or both of —NHgroup and —NH₂ group.

The invention's discovery of the invention's compositions that contain acombination of an acid and organic diol was premised, in part, on thesurprising discovery of synergistic antimicrobial activity of thiscombination, including at acidic pH.

The invention's discovery of the invention's compositions that contain acombination of an acid, organic diol and an oxidizer was premised, inpart, on the surprising discovery that adding an organic diol to anoxidizer in solution resulted in the ability to alter (increase ordecrease) the pH of the solution independently of the bio-load, whilealso providing synergistic antimicrobial activity of the acid, organicdiol and oxidizer.

In addition, inclusion of the organic diol unexpectedly reduced theundesirable odor when an oxidizer is included in the composition,particularly in the presence of bioload. This allowed, in oneembodiment, the further addition of acids to the solution, whichresulted in surprising synergistic antimicrobial activity by the acid,organic diol, and oxidizing agent. Furthermore, the inventor made thesurprising observation that particular embodiments of the invention'scompositions were capable of 100% reduction in microbial contaminationat the chilling temperatures (e.g., from 30° F. to 40° F.) that areroutinely used during standard processing steps for agriculturalproducts, while not resulting in adverse effects on the odor, texture,color and/or appearance of the agricultural products. A furthersurprising aspect of the invention's compositions is that they had abroad spectrum of antimicrobial activity against bacterial,bacteriophage, viral, fungal and nematode microbes, using a wide varietyof agricultural products and inanimate objects. Moreover, theinvention's compositions surprisingly reduced discoloration (e.g.,pinking and browning) of agricultural products. Yet another surprisingproperty of the invention's compositions is that they showed adifferential effect on pathogenic and non-pathogenic microbes, byreducing the number of pathogenic bacteria to a greater extent than thereduction of non-pathogenic bacteria and of non-pathogenicbacteriophage.

The invention's compositions have several additional advantages overantimicrobial compositions of the prior art. For example, in oneembodiment, the invention's compositions are clear solutions,non-foaming during use, and continue to exhibit residual antimicrobialactivity after drying.

In addition, the invention's compositions in one embodiment exhibitantimicrobial activity at acidic pH, thus advantageously allowing, inone embodiment, chlorine salts to remain in solution.

Moreover, after drying, the invention's compositions form avery thinfilm on the surface (including cracks and open stomata) of the treatedproducts consisting of the composition's components. This film is ediblebecause of its low toxicity to higher life forms, while continuing toexhibit antimicrobial activity, thus eliminating the need, andenvironmental and monetary cost, for rinsing off the invention'scomposition from the treated products (e.g., agricultural products).Moreover, a film can be placed on machines (or portions thereof such ascutting blades) that process agricultural products.

In addition, in one embodiment, the invention's compositions may beapplied pre-harvest and post harvest of agricultural products, thusadvantageously extending the period of exposure to antimicrobialactivity, and resulting in increased shelf life.

Another advantage of the invention's compositions is that theirantimicrobial activity is resistant to particulate loads, increasedturbidity, and plant fluids. They also decrease turbidity in thesolution, allow for lower processing temperatures (one to two ° F.), anddecrease the incidence of ice crystal formation on the treated materialsat refrigerated temperatures, thereby preserving surface quality.

A further advantage is that the invention's compositions arenon-irritating when applied to skin as antiseptic even after 12 hours ofcontact, thus making them useful in medical applications. In oneembodiment, compositions of the present invention can be applieddirectly to the skin. In another embodiment, the present inventioncontemplates surgical dressings, bandages and surgical tapes comprisingone or more of the compositions set forth herein (see Tables below).Illustrative examples of such dressings and tapes include, but are notlimited to, sheets of material, surgical swabs, gauze pads, closurestrips, compress bandages, surgical tape, and the like. In a furtherembodiment, medical devices are contacted (and preferably coated, e.g.,coated with a film) with compositions set forth herein (see Tablesbelow). Such devices can be used both outside and in the body, includingimplantable medical devices (e.g., staples, shunts, stents, stitches,tubing, and the like). In one embodiment, teeth or dental products arecontacted with compositions set forth herein. In yet another embodiment,contact lenses and related eye care articles are contacted withcompositions set forth herein.

An additional advantage is that the invention's compositions may be madeas concentrates and used in situ and ex situ. Yet another advantage isthat they may be readily applied by standard methods such as spraying,dipping, wiping, etc., thus avoiding the need for costly modificationsto existing commercial applications.

Thus, the invention's compositions are useful for antimicrobialapplications in a variety of settings, such as before harvesting and/orat the point of harvesting and/or after harvesting of agriculturalproducts (including food products, ornamental plants, etc.), at foodproduction facilities including the disinfection of food processingequipment and harvesting equipment, in hospitals, offices, residences,etc. For example, the invention's compositions may be used pre-harvestfor microbe reduction on living plants. Also, ice made of theinvention's compositions may be used for cold storage of shellfish,fruits, vegetables and meats. The compositions also may be used toreduce undesirable odors from bacterial breakdown of food materials.Further utilities include reduction of microbial infection of skin, suchas athlete's foot fungus “tinea pedis” (Trichophyton).

In addition, the invention's compositions may be used for rendering anytype of surface resistant to mold, for cleaning and/or pre-treating suchsurfaces, and for rendering surfaces contaminated with potentially oractually hazardous microbes such as, but not limited to, mold spores,bacteria, viruses, protozoa and/or any biological warfare agents, safeto handle and/or safe to dispose of.

Additional advantages, surprising results and attributes of theinvention are further described below under (A) Compositions and (B)Methods.

A. Compositions

The invention provides antimicrobial compositions comprising one or moreacid and one or more organic diol. In one embodiment, the invention'scompositions have an acidic pH. The compositions may optionally furthercontain one or more oxidizing agent (including stabilized oxidizingagent and/or unstabilized oxidizing agent), and/or one or moresurfactant. In particular embodiments, the acid lacks one or both of —NHgroup and-NH₂ group.

The invention's compositions are exemplified by, but not limited to,compositions AA, BB, CC, DD, A, B, C, D, E, F, G, H, I, J, K, L, M, Nand O in Table 1, which are illustrated by compositions xx1, xx2, xx3,xx4, x1a, x1, x2, x3, x4, x5, x6, x7, x8, x9, x10, x11, x12, x13, x14and x15 in Table 2 and by compositions a1 to a5 in Tables 5 and 8.

TABLE 1 Exemplary compositions of the invention⁽¹⁾ Acid Diol OxidizingAgent Surfactant Ref.⁽²⁾ (wgt. %) (wgt. %) (wgt. %) (wgt. %) AA0.001-0.9 0.001-2.0 ortho-Phosphoric acid Propylene glycol BB⁽⁷⁾0.001-1.0 0.001-2.0 ortho-Phosphoric acid 1,3-Butanediol CC⁽⁸⁾ 0.003-5.00.001-2.5 Hydrochloric acid Propylene glycol DD^((7,8)) 0.003-5.00.001-2.0 Hydrochloric acid 1,3-Butanediol A⁽³⁾ 0.001-0.9 0.001-2.00.005-2.9 ortho-Phosphoric acid Propylene glycol Hypochlorous acid B0.001-0.9 0.001-3.0 0.005-2.9 ortho-Phosphoric acid Propylene glycolHypochlorous acid and 0.05-0.9 Bromine C⁽⁴⁾ 20-30 3 20 ortho-Phosphoricacid Propylene glycol Hydrogen peroxide D⁽⁵⁾ 30 3 20 0.01-1.9ortho-Phosphoric acid Propylene glycol Hydrogen peroxide Sodium laurylsulfate E 0.001-0.9 0.01-3.0 0.05-0.9 0.01-1.9 ortho-Phosphoric acidPropylene glycol Hypochlorous acid Sodium lauryl sulfate and 0.05-0.9Bromine F 0.001-0.05 0.01-3.0 0.05-3.9 ortho-Phosphoric acid Propyleneglycol Peroxyacetic acid G 0.001-0.05 0.01-3.0 0.05-3.9 0.01-1.9ortho-Phosphoric acid Propylene glycol Peroxyacetic acid Octyl phenolethoxylate H 0.001-0.05 0.01-3.0 0.0003-1.9 ortho-Phosphoric acidPropylene glycol Ozone I 0.001-0.05 0.01-3.0 0.0003-1.9 0.01-1.9ortho-Phosphoric acid Propylene glycol Ozone Octyl phenol ethoxylate J1-6 0.1-0.6 0.7-4 ortho-Phosphoric acid Propylene glycol Hydrogenperoxide K⁽⁶⁾ 30 3 20 Less than 0.1 wgt. % ortho-Phosphoric acidPropylene glycol Hydrogen peroxide of a surfactant to the concentratewhen it is to be applied to surfaces L 0.3-5.0 0.1-0.6 0.7-4Hydrochloric acid Propylene glycol Hydrogen peroxide M⁽⁶⁾ 20 3 20 Withor without Hydrochloric acid Propylene glycol Hydrogen peroxide 0.1 wgt.% of a surfactant N 0.9-8 0.1-0.6 0.7-4 Citric acid Propylene glycolHydrogen peroxide O⁽⁶⁾ 35 3 20 Addition of less than Citric acidPropylene glycol Hydrogen peroxide 0.1 wgt. % of a surfactant to theconcentrate when it is to be applied to surfaces. ⁽¹⁾Water was used assolvent. ⁽²⁾pH of compositions other than concentrated compositions, isacidic. ⁽³⁾Concentrated compositions, such as compositions C, D, K, M,and O are diluted to a final concentration having an acidic pH less thanpH 7.0. ⁽⁴⁾Composition A is food-safe and does not need to be rinsedoff. ⁽⁵⁾Composition C is a concentrated antimicrobial composition thathas antimicrobial effects when diluted to about one hundred to one inpotable water, is food-safe and does not need to be rinsed off.⁽⁶⁾Composition D is a concentrated antimicrobial composition that hasantimicrobial effects when diluted to about one hundred to one inpotable water, is food-safe and does not need to be rinsed off.⁽⁷⁾Compositions K, M and O are concentrated antimicrobial compositions.⁽⁸⁾With higher concentrations of 1,3-butanediol, the treated materialhad a slight odor. ⁽⁹⁾Compositions CC and DD killed Lactobacillus, andbroad leaf material bleached quicker than with compositions AA and BB.The exemplary compositions in Table 1 above are illustrated bycompositions xx1, xx2, xx3, xx4, x1a, x1, x2, x3, x4, x5, x6, x7, x8,x9, x10, x11, x12, x13, x14 and x15 in Table 2 below (See also Tables 5and 8).

TABLE 2 Additional Exemplary compositions of the invention. OxidizingSub Acid Diol Agent Surfactant Ref.⁽¹⁾ Ref.⁽²⁾ (wgt. %) (wgt. %) (wgt.%) (wgt. %) pH⁽⁹⁾ AA xx1 0.035 0.02 ortho-Phosphoric acid Propyleneglycol BB xx2 0.03 0.012 ortho-Phosphoric acid 1,3- Butanediol CC xx30.01 0.018 Hydrochloric acid Propylene glycol DD xx4 0.008 0.005Hydrochloric acid 1,3- Butanediol A x1a 0.03 0.02 0.005 4ortho-Phosphoric acid Propylene Hypochlorous glycol acid A x1 0.08 0.090.05   3-4.1 ortho-Phosphoric acid Propylene Hypochlorous glycol acid Bx2 0.08 0.09 0.01   4-5.5 ortho-Phosphoric acid Propylene Hypochlorousglycol acid and 0.05 Bromine C⁽³⁾ x3 21 3 20 0.5-1   ortho-Phosphoricacid Propylene Hydrogen glycol peroxide D⁽⁴⁾ x4 30 3 20 0.01 0.5-1.5ortho-Phosphoric acid Propylene Hydrogen Sodium lauryl glycol peroxidesulfate E⁽⁵⁾ x5 0.07 0.1 0.007 0.03 3.3-4.5 ortho-Phosphoric acidPropylene Hypochlorous Sodium lauryl glycol acid sulfate and 0.08Bromine F x6 0.003 0.1 0.5 3-4 ortho-Phosphoric acid PropylenePeroxyacetic glycol acid G x7 0.003 0.14 0.5 0.015 3-4 ortho-Phosphoricacid Propylene Peroxyacetic octyl phenol glycol acid ethoxylate H x80.02 0.1 0.001 5-6 ortho-Phosphoric acid Propylene Ozone glycol I x90.02 0.12 0.001 0.015 5.5-6.5 ortho-Phosphoric acid Propylene Ozoneoctyl phenol glycol ethoxylate J x10 1 0.1 2.1 less thanortho-Phosphoric acid Propylene Hydrogen 2.0 glycol peroxide K⁽⁶⁾ x11 303 20 0.001 — ortho-Phosphoric acid Propylene Hydrogen Cetylpyridiniumglycol peroxide chloride L x12 0.3 0.1 3 less than Hydrochloric acidPropylene Hydrogen 2.0 glycol peroxide 2.0 M⁽⁷⁾ x13 20 3 20 —Hydrochloric acid Propylene Hydrogen glycol peroxide N x14 1 0.3 2 lessthan Citric acid Propylene Hydrogen 2.0 glycol peroxide O⁽⁸⁾ x15 35 3 200.001 — Citric acid Propylene Hydrogen sodium lauryl glycol peroxidesulfate ⁽¹⁾Composition reference letter corresponds to the samereference letter in Table 1. ⁽²⁾Composition sub-reference lettering(e.g., x1) refers to an exemplary composition within the range ofconcentrations of the reference composition (e.g., composition A).⁽³⁾Composition C-x3 is a concentrated antimicrobial composition.⁽⁴⁾Composition D-x4 is a concentrated antimicrobial composition, and ispreferably prepared by first mixing sodium lauryl sulfate with propyleneglycol. ⁽⁵⁾Composition F-x5 is preferably prepared by first mixingsodium lauryl sulfate with propylene glycol. ⁽⁶⁾Composition K-x11 is aconcentrated antimicrobial composition. ⁽⁷⁾Composition M-x13 is aconcentrated antimicrobial composition. ⁽⁸⁾Composition O-x15 is aconcentrated antimicrobial composition. ⁽⁹⁾pH at 34° F. Concentratedcompositions are diluted to a working concentration having an acidic pHless than 7.0.

The antimicrobial activity of the exemplary compositions x1 to x15 isshown in Table 6 on bacterial, bacteriophage, viral, fungal and nematodemicrobes, using a wide variety of agricultural products (including leafyvegetables, fruiting and flowering vegetables, podded vegetables, bulband stem vegetables, root and tuberous vegetables, Rosaceae familyfruits, Bramble fruits, berries, Asian fruit, North America fruit, rawmeat, raw poultry, raw seafood, fresh seeds and nuts, eggs, fresh herbs,dried herbs, animal hides, feathers) and inanimate objects (such asthose encountered in hospitals, food processing plants, residentialbuildings, office buildings, etc.). In addition, the antimicrobialactivity of the exemplary compositions a1 to a5 is shown in Tables 5 and8 on the highly pathogenic E. coli.

The components of the invention's compositions include those that areapproved and not approved by the Food and Drug Administration (FDA) forone or more uses, including, but not limited to, compounds that aregenerally recognized as safe (GRAS) and/or food grade compounds (FCC)and/or registered with the Environmental Protection Agency (EPA).

The terms “generally recognized as safe” and “GRAS” when in reference toa compound refer to each of the substances in 21 C.F.R., §182 and §184,approved before 1958 by the FDA and/or USDA, items published in theFederal Register, as well as other compounds (even if not on the GRASlist) that are accepted in the industry as safe for one or more uses byan animal, including by a human. As used herein, “food grade” and “FCC”refer to the Food Chemical Codex (FCC) that serves as a guide for foodgrade quality consistent with good manufacturing practices and ispublished by U.S. Pharmacopeia (USP). For example, and withoutlimitation, organic diols approved by the FDA and within the scope ofthe invention include propylene glycol (1,2-Propanediol),1,3-butanediol, 1,2-ocatnediol, 1,5-pentanediol, and polyethyleneglycol. Organic diols within the scope of the invention that are not yetapproved by the FDA include ethylene glycol and 1,7-heptanediol.

The invention's compositions may contain one or more acid. An “acid” isa chemical compound that, when dissolved in water, gives a solution witha hydrogen ion activity greater than in pure water, i.e. has an acidicpH. Acids may be organic or inorganic. Organic acids are acids thatcontain carbon. Inorganic acids, also referred to as mineral acids, areacids that do not contain carbon. The Chemical Abstracts Service(“CAS”), Columbus, Ohio, USA, provides a comprehensive repository fordata on organic and inorganic acids. Organic acids include, but are notlimited to, hydroxyethanoic acid (CAS 79-14-1), citric acid (CAS5949-29-1), ascorbic acid (CAS 50-81-7), lactic acid (CAS 50-21-5),malic acid (CAS 617-48-1), octenic acid (CAS 1871-67-6), oxalic acid(CAS 144-62-7), ursolic acid (CAS 77-52-1), and salts thereof Inorganicacids include, but are not limited to, ortho-phosphoric acid (CAS7664-38-2), chromic acid (CAS 1308-38-9), hydrobromic acid (CAS10035-10-6), hydrochloric acid (also known as muratic acid) (CAS7647-01-0), nitric acid (CAS 7697-37-2), sulfuric acid (CAS 7664-93-9)and salts thereof Other acids contemplated within the scope of theinvention include, without limitation, acetic acid, adipic acid, benzoicacid, glutaric acid, isoascorbic acid, mandelic acid, propionic acid,salicylic acid, sorbic acid, succinic acid, tartaric acid (Koefod U.S.Pat. No. 7,090,882), sodium acid pyrophosphate, acidic sodiumhexametaphosphate, ethylenediaminetetraacetic acid and salts thereof(Andrews et al., U.S. Pat. No. 5,490,992). Because of their lowsolubility in water, octenic acid and ursolic acid may first bedissolved in an organic solvent (e.g., glycol, diol, ethanol at atemperature equal to or greater than 100° F., etc.) before mixing withwater. Nitric acid at some concentrations may be toxic to humans, whichmakes its use on foodstuffs less desirable than other acids. Oxalic acidand chromic acid at some concentrations react with some materials, suchas steel, so their use in cleaning equipment may be less desirable thanother acids. In one embodiment, the acid lacks —NH groups and/or —NH₂groups (e.g., is not a sulfamic acid).

In some embodiments, the amount of acid in the invention's concentratedcompositions may be from 2 to 50 wgt. %, preferably from 10 to 35 wgt.%, and more preferably from 20 to 30 wgt. %. In particular embodiments,the concentrate is typically diluted with water to provide an acidconcentration of from 0.001 to 2 wgt. %, preferably from 0.003 to 1 wgt.%, and more preferably from 0.01 to 0.10 wgt. %.

The invention's compositions may contain one or more organic diol. A“diol” or “glycol” is a chemical compound containing two hydroxyl (—OH)groups. A diol may be a vicinal diol or a geminal diol, including analiphatic diol. A vicinal diol has hydroxyl groups bonded to adjacentatoms, e.g., ethylene glycol and propylene glycol. A geminal diol hashydroxyl groups bonded to the same atom, e.g., methanediol (H₂C(OH)₂),1,3-butanediol, bisphenol A, etc. Organic diols may contain a branchedchain or an unbranched chain of carbon atoms. Organic diols arepreferably aliphatic diols (i.e., an organic diol composed of carbon andhydrogn and does not contain an aromatic ring), and more preferablysimple aliphatic diols. A “simple aliphatic diol” is an aliphatic diolin which all the carbon atoms are covalently bonded to only hydrogen,and is exemplified by ethylene glycol, propylene glycol, 1,3-butanediol,etc. In contrast, a “complex aliphatic diol” is an aliphatic diol inwhich at least one carbon atom is covalently bonded to one or more atomsother than hydrogen. Thus, a complex aliphatic diol may contain —COOH,—NH₂, etc. In some embodiments, organic diols may contain one or moreether group (glycol ether) and/or one or more ester group (glycolester), including monoesters and diesters. In another embodiment,organic diols may have one hydroxyl group esterified with a saturatedand/or unsaturated aliphatic acid other than benzoic acid, and the otherhydroxyl group esterified with the same or different saturated and/orunsaturated aliphatic acid other than benzoic acid. Exemplary organicdiols include, without limitation, propylene glycol (CAS 57-55-6),ethylene glycol (CAS 107-21-1), 1,3-butanediol (CAS 107-88-0),1,7-heptanediol (CAS 629-30-1), 1,2-octanediol (CAS 1117-86-8),1,5-pentanediol (CAS 111-29-5) and polyethylene glycol (CAS 25322-68-3).In some embodiments, organic diols contain from 2 to 20, such as from 2to 10, and/or such as from 2 to 5 carbon atoms. For example, the diolethylene glycol contains 2 carbon atoms, 1,3-butanediol contains 4carbon atoms, 1,5-pentanediol contains 5 carbon atoms, 1,7-heptanediolcontains 7 carbon atoms, and 1,2-octanediol contains 8 carbon atoms.

In some embodiments, the amount of organic diol in the invention'sconcentrated compositions may be from 2 to 5 wgt. %, preferably from 3to 4 wgt. %, and more preferably 3 wgt. %. In particular embodiments,the concentrate is typically diluted with water to provide an organicdiol concentration of from 0.01 to 1 wgt. %, preferably from 0.09 to 0.3wgt. %, and more preferably from 0.09 to 0.15 wgt. %.

The invention's compositions may contain one or more oxidizing agent.“Oxidizing agent,” “oxidant” and “oxidizer” are interchangeably used torefer to a chemical compound that transfers oxygen atoms or gainselectrons in a redox chemical reaction. In both cases, the oxidizingagent becomes reduced in the process. Exemplary oxidizing agentsinclude, without limitation, bromine (CAS 7726-95-6), hypochlorous acid(CAS 7790-92-3, also referred to as IUPAC name chloric(I) acid),hydrogen peroxide (CAS 7722-84-1), peroxyacetic acid (PAA) (CAS79-21-0), and ozone (CAS 10028-15-6). Hypochlorous acid cannot beisolated in pure form, but can be derived from, without limitation,sodium hyporchlorite, calcium hyporchlorite, chlorine dioxide andchlorine. Further oxidizing agents include sodium bromide, peraceticacid, chlorine dioxide, sodium hypochlorite, hydrogen peroxide (BautistaU.S. Pat. No. 6,045,846) and ozone. Ozone may be introduced into a fluidusing methods known in the art (Conners et al., U.S. Pat. No.6,086,833).

In preferred embodiments, the invention's compositions contain more thanone oxidizer to produce a greater reduction in antimicrobial activityand/or in exposure times when compared to single oxidizers in order toobtain the same antimicrobial activity.

In some embodiments, the amount of oxidizer in the invention'sconcentrated compositions may be from 2 to 30 wgt. %, preferably from 10to 30 wgt. %, and more preferably from 10 to 20 wgt. %. In particularembodiments, the concentrate is typically diluted with water to providean oxidizer concentration of from 0.01 to 5 wgt. %, preferably from0.001 to 3 wgt. %, and more preferably from 0.01 to 1 wgt. %. Forexample, levels of hypochlorous acid and hypochlorite are reported asCl₂, using the standard DPD (N,N-diethyl-p-phenylenediamine)colorimetric test method (Hach Chemical Co., Loveland Colo.), describedin Harp, U.S. Pat. No. 5,362,650 and Kroll, U.S. Pat. No. 6,180,412, byemploying standard instruments, such as model number 5870000 pocketcolorimeter II “2.”

In contrast to the art (e.g., Williams, U.S. Patent Application20060003023), in particular embodiments, the invention's compositionscontain an “unstabilized oxidizer” (“USOX”), i.e., contain an oxidizingagent in the absence of an oxidizing agent stabilizer. A “compound thatstabilizes an oxidizing agent” and “oxidizing agent stabilizer”interchangeably refer to a compound that reduces the rate of decrease inconcentration of an oxidizing agent, such as that resulting fromdegradation of the oxidizer. Oxidizing agent stabilizers include, forexample, sulfamic acid, alkali metal sulfamate (Williams, U.S. PatentApplication 20060003023), amide derivatives of carbonic acid, hydrogencyanide, carboxylic acid, amino acid, sulfuric acid, phosphoric acid,boric acid, urea, thiourea, creatinines, cyanuric acids, alkylhydantoins, mono-ethanolamine, di-ethanolamine, organic sulfonamides,biuret, sulfamic acid and salts thereof; organic sulfamic acid, melamine(Shim et al., U.S. Pat. Nos. 6,478,972 and 6,533,958).

Oxidizers are unstable, and their rate of oxidation may change (i.e.,increase or decrease) and may become uncontrolled, resulting indepletion of the oxidizing agent in a short period of time (such as lessthan 6 hours, Example 9). To avoid this problem, the prior art, such asSelf et al., U.S. Pat. No. 3,328,294, discloses a stabilized oxidizer(“SOX”) produced by reacting the unstabilized oxidizer NaOBr with anequal molar ratio of the stabilizer sulfamic acid. They disclose thatthe reaction does not go un-controlled and the stabilized oxidizer maylinger for days. Because of these characteristics they caution againstthe use of their compositions on foods and in potable water.

In contrast to the prior art, particular embodiments, the invention usesunstabilized oxidizers in order to take advantage of newer technologyand increase the log microbial kill rate. It is the inventor's view thatunstabilized oxidizers provide an instantaneous reaction in a processstream or on contacting a surface, a reduction in toxicity to humans andanimals, an almost immediate kill of contacted microbes, shorterprocessing time, instantaneous concentration control at product contactpoints, and take advantage of new control systems for fast automaticcontrol of active ingredients.

The invention's compositions may contain one or more surfactant. A“surfactant” is a chemical compound that lowers the surface tension of aliquid, allowing easier spreading of the liquid, and thereby acting as awetting agent. In one embodiment, a surfactant is an organic compoundthat is amphiphilic, meaning that it contains one or more hydrophobicgroups (“tails”) and one or more hydrophilic groups (“heads”).Surfactants may be anionic, neutral or cationic. “Anionic surfactants”are surfactants that dissolve in water to release an anion, and include,for example, sodium lauryl ether sulfate, also referred to as sodiumlauryl sulfate (CAS 009004-82-4), ammonium lauryl sulfate (CAS2235-54-3), alkylbenzene sulfonic acid (CAS 27176-87-0), sodium2-ethylhexyl sulfate (CAS 126-92-1), and dioctyl sodium sulfosuccinate(Andrews et al., U.S. Pat. No. 5,490,992). “Neutral surfactants” aresurfactants that dissolve in water without releasing an anion or cation,and include, without limitation, octyl phenol ethoxylate (CAS9002-93-1), glyceryl monostearate (CAS 31566-31-1), polyglyceryl-10decaoleate (CAS 011094-60-3), and lauryl lactyl lactate (CAS910661-93-7). “Cationic surfactants” are surfactants that dissolve inwater to release a cation, and include the exemplary cetrimonium bromide(CAS 57-09-0), cetylpyridinium chloride (CAS 123-03-5), benzalkoniumchloride (CAS 8001-54-5), and cocamidopropyl betaine (CAS 86438-79-1).Additional examples of surfactants include those that contain ethyleneoxide moieties and/or propylene oxide moieties. Yet more examples ofsurfactants include linear alkylbenzene sulfonates, alcohol sulfates,alpha-olefin sulfonates, alcohol ethoxylates, nonylphenyl ethoxylates,alkylpolyglucosides, fatty alkanoamides, fatty amine oxides, sodiumdioctylsulfosuccinate, dodecylbenzene sulfonic acid and salts thereof,the sodium salt of sulfonated oleic acid, sodium dodecylbenzenesulfonate, dodecyidiphenyloxidedisulfonic acid and salts thereof (Koefodet al., U.S. Pat. No. 7,090,882, Andrews et al., U.S. Pat. No.5,490,992).

In some embodiments, the amount of surfactant in the invention'sconcentrated compositions may be from 0.05 to 0.5 wgt. %, preferablyfrom 0.01 to 0.03 wgt. %, and more preferably from 0.01 to 0.15 wgt. %.In particular embodiments, the concentrate is typically diluted withwater to provide a surfactant concentration of from 0.01 to 0.05 wgt. %,preferably from 0.001 to 0.005 wgt. %, and more preferably from 0.001wgt. % to 0.03 wgt. %.

The invention additionally provides compositions comprising aconcentrated solution of any of the compositions disclosed herein. A“concentrate” of a first composition refers to a compositions thatcontains the same components and the same ratio of these components(relative to each other) as in the first composition, wherein the amountof each component in the first composition is multiplied by the samefold factor to yield the amount of that component in the concentratedcomposition. The fold factor may be from 2 to 100,000, from 5 to 10,000,from 10 to 1,000, and from 100 to 500. For example, for a compositioncontaining 4 wgt. % component A and 6 wgt. % component B, a 2-foldconcentrate of this composition contains 8 wgt. % component A and 12wgt. % component B, and a 3-fold concentrate of this compositioncontains 12 wgt. % component A and 18 wgt. % component B. Concentratesmay be used directly or diluted. They provide ease of handling andshipping (because of smaller volumes). Exemplary concentrates arecompositions C, D, J, K, M, O in Table 1, as exemplified by compositionsx3, x10, x11, x13 and x15 of Table 2.

The invention also contemplates compositions comprising dilutedsolutions of any of the compositions disclosed herein. A “diluted”solution of a first composition refers to a compositions that containsthe same components and the same ratio of these components (relative toeach other) as in the first composition, wherein the amount of eachcomponent in the first composition is divided by the same fold factor toyield the amount of that component in the concentrated composition. Thefold factor may be from 2 to 100,000, from 5 to 10,000, from 10 to1,000, and from 100 to 500. For example, for a composition containing 4wgt. % component A and 6 wgt. % component B, a 2-fold dilution of thiscomposition contains 2 wgt. % component A and 3 wgt. % component B.Diluted solutions may be used directly or further diluted. Exemplarydiluted solutions are compositions A, B,E, F, G, H, I, L, N, O in Table1, as exemplified by compositions xx1, xx2, xx3, xx4, x1a, x1, x2, x4,x5, x6, x7, x8, x9, x12, x14 and in Table 2 and by compositions a1 to a5of Tables 5 and 8.

The invention's compositions may be prepared by adding and mixing theacid alone into water, followed by adding and mixing in the diol, andthen adding and mixing in the oxidizing agent. Surfactant may besubsequently added and mixed in. The compositions may be compounded attemperatures between the freezing point and the boiling point of thesolvent (e.g., water) used.

When making concentrates, the diol and acid may be mixed together inconcentrated quantities, followed by the surfactant, if desired, fordelivery to a mixture of oxidizer and solvent (e.g., water).Alternatively, the diol and oxidizer may be mixed together inconcentrated quantities for delivery to a mixture of acid and solvent(e.g., water). Concentrates may be stored for more than 1 week. Theinvention's compositions may be used in a variety of ways as furtherdescribed below.

B. Methods

In one embodiment, the invention's compositions have antimicrobialactivity and may be used in methods for reducing the number of microbeson a surface, comprising (a) providing (i) a first surface comprisingmicrobes, (ii) an antimicrobially effective amount of any of theinvention's compositions, and b) contacting the surface with thecomposition under conditions that produce a contacted surface comprisinga reduced number of the microbes compared to the number of the microbeson the first surface in the absence of the contacting step.

The term “antimicrobial” and “antimicrobial activity” when in referenceto a compound refers to a compound that reduces the number of and/orrate of growth of a microbe compared to the number and/or rate of growthof the microbe in the absence of the compound. In one embodiment, thenumber of and/or rate of growth of a microbe in the presence of anantimicrobial compound is at least 10% lower than, at least 25% lowerthan, at least 50% lower than, at least 75% lower than, at least 90%lower than, at least 99%, and/or at least 100% lower than the number ofand/or rate of growth of the microbe in the absence of the antimicrobialcompound. In another embodiment, a 9-log number of microbe is reduced byat least 3-log, at least 5-log, at least 7-log, and/or at least 9-log inthe presence of the antimicrobial compound.

An “antimicrobially effective amount” of a compound or compositionrefers to an amount of the compound or composition that hasantimicrobial activity, including microbistatic amounts and microbicidalamounts.

“MIC” and “minimum inhibitory concentration” refer to the lowestconcentration that is microbistatic under a specific set of conditions.

“MBC” and “minimum biocidal concentration” refer to the lowestconcentration that is microbicidal under a specific set of conditions.

An antimicrobial can be antibacterial, antifungal, antiviral and/orantinematode. An antimicrobial can be microbistatic, microbicidal, orboth. An antimicrobial is “microbistatic” (e.g., bacteriostatic,fungistatic, etc.) if it reduces cell division by an amount less than100%, without or without reducing cell viability. An antimicrobial is“microbicidal” (e.g., bactericidal, fungicidal, etc.) if it reduces cellviability by 100%, i.e., causes 100% cell death. Cell death is commonlydetected by the absence of cell growth in liquid growth medium (e.g.,absence of turbidity) or on a solid surface (e.g., absence of colonyformation on agar). A “sterilizer,” “sanitizer” and “disinfectant” aremicrobicidal. In contrast, a “preservative” is microbistatic. Certainmicrobistatic compositions are not bactericidal at any concentration.

Those of skill in the art know that a composition may have microbistaticor microbicidal activity by altering the concentration of itscomponents, temperature, and contact time with a surface. For example, acomposition that is microbistatic at a given concentration may bemicrobicidal at a higher concentration. Also, a composition that ismicrobistatic at a given temperature may be microbicidal at a highertemperature. Similarly, a composition that is microbistatic at aparticular contact time may be microbicidal at a longer contact timewith a surface.

Methods for determining antimicrobial activity are known in the art,e.g., Andrews et al., U.S. Pat. No. 5,490,992, Bailey et al., U.S. Pat.No. 4,107,192and disclosed herein. For example, one assay involvesexposing a bacterial strain (e.g., Escherichia coli) to a testcomposition on a particular substrate (e.g., an agricultural product) ata predetermined bacterial level in a culture media at an appropriatetemperature. After a sufficient amount of contact time, an aliquot of asample containing the exposed bacteria is collected, diluted and platedon agar. The plated sample of bacteria is then incubated for about fortyeight (48) hours and the number of viable bacterial colonies growing onagar is counted. Once colonies have been counted the reduction in thenumber of bacteria caused by the test composition is determined.Reduction in bacteria is typically reported as the difference betweenthe log₁₀ of the initial inoculant's count and the log₁₀ of theinoculant's count after exposure to the test composition. Assays mayalso be subjective, such as by visually observing a difference in theamount of spoilage of agricultural products.

The invention's compositions exhibit surprising synergistic activitywith respect to, for example, antimicrobial activity of theircomponents. “Synergistic” activity of a combination of two or morecomponents means that the activity of the combination of two or morecomponents is greater than the activity of each component alone. Withoutintending to limit the invention to any particular method ofcalculation, in one embodiment, synergy may be determined using theindustrially accepted method described by Kul et al., AppliedMicrobiology 9:538-541 (1961) (see also Williams, U.S. PatentApplication 20060003023), using the ratio determined by the formula:

Q _(a) /Q _(A) +Q _(b) /Q _(B)=Synergy Index (“SP”) wherein:

Q_(A)=concentration of compound A (first component) in ppm, actingalone, which produced an end point (MIC or MBC of Compound A).

Q_(a)=concentration of compound A in ppm, in the mixture, which producedan end point.

Q_(B)=concentration of compound B (second component) in ppm, actingalone, which produced an end point (MIC or MBC of Compound B).

Q_(b)=concentration of compound B in ppm, in the mixture, which producedan end point.

When the sum of Q_(a)/Q_(A) and Q_(b)/Q_(B) is greater than one,antagonism is indicated. When the sum is equal to one, additivity isindicated, and when less than one, synergism is demonstrated. The lowerthe SI, the greater the synergy shown by that particular mixture.

Thus, in one embodiment, synergy of the combination of acid and organicdiol is obtained where the amount of each of the acid alone and of theorganic diol alone has lower antimicrobial activity compared to theantimicrobial activity of a combination of the acid and the organicdiol. In another embodiment, synergy of the combination of acid, organicdiol and oxidizing agent is obtained where the amount of each of (a) theacid alone, (b) the organic diol alone, (c) the oxidizing agent alone,(d) combination of the acid and the organic diol, (e) combination of theacid and the oxidizing agent, and (f) combination of the organic dioland the oxidizing agent, has lower antimicrobial activity compared tothe antimicrobial activity of a combination of the acid, the organicdiol and the oxidizing agent.

For example, Table 3A shows antimicrobial synergy of propylene glycoland phosphoric acid, synergy of propylene glycol, phosphoric acid andhydrogen peroxide, and synergy of propylene glycol, phosphoric acid,hydrogen peroxide and sodium lauryl sulfate.

Also, Table 4 shows antimicrobial synergy of two components whencompared to single components of Table 3A.

In another example, data herein shows synergistic antimicrobial activityof the components of composition “a1” that contains hypochlorous acid0.01 wgt. %, phosphoric acid 1 wgt. %, and propylene glycol 2 wgt. %.

In particular, Table 3B shows that 180 minute treatment of spinach witheach of hypochlorous acid 0.01 wgt. %, phosphoric acid 1 wgt. % andpropylene glycol 2 wgt. % alone in water causes a 5.01-log, 4.13-log,and 3.98-log reduction, respectively, in a 6.87-log E. coli inoculum.Table 4 shows similarly low antimicrobial activity by a combination oftwo components of hypochlorous acid 0.01 wgt. % and phosphoric acid 1wgt. %, which resulted in a 4.9-log reduction in a 6.87-log E. coliinoculum. In contrast, Table 5 shows that 180 minute treatment ofspinach with the combination “a1” that contained hypochlorous acid 0.01wgt. %, phosphoric acid 1 wgt. %, and propylene glycol 2 wgt. % resultedin a 6.57-log reduction in a 6.57-log E. coli inoculum.

Similarly, Table 3B shows that 180 minute treatment of spinach with eachof bromine 0.08 wgt. %, phosphoric acid 1 wgt. % and propylene glycol 2wgt. % alone caused a 4.44-log, 4.13-log, and 3.98-log reduction,respectively, in a 6.87-log E. coli inoculum. Table 4 shows similarlylow antimicrobial activity by a combination of two components of bromine0.08 wgt. % and phosphoric acid 1 wgt. % which resulted in a 4.88-logreduction in a 6.87-log E. coli inoculum. In contrast, Table 5 showsthat 180 minute treatment of spinach with the combination “a2” thatcontained bromine 0.08 wgt. %, phosphoric acid 1 wgt. % and propyleneglycol 2 wgt. % resulted in a 6.49-log reduction in a 6.57-log E. coliinoculum.

In yet another example, Table 3B shows that 180 minute treatment ofspinach with each of hypochlorous acid 0.01 wgt. %, bromine 0.05 wgt. %,phosphoric acid 1 wgt. %, and propylene glycol 2 wgt. % alone in watercaused a 5.01-log, 4.44-log, 4.13-log and 3.98-log reduction,respectively, in a 6.87-log E. coli inoculum. Table 4 shows similarlylow antimicrobial activity by a combination of two components of bromine0.08 wgt. % and phosphoric acid 1 wgt. % which resulted in a 4.88-logreduction in a 6.87-log E. coli inoculum. In contrast, Table 5 showsthat 180 minute treatment of spinach with the combination “a5” thatcontained hypochlorous acid 0.01 wgt. %, bromine 0.05 wgt. %, phosphoricacid 1 wgt. %, and propylene glycol 2 wgt. %, resulted in a 6.57-logreduction in a 6.57-log E. coli inoculum.

The invention's compositions showed very effective antimicrobialactivity (Tables 5, 6, 8 and 9). For example, Table 6 shows theantimicrobial activity of the invention's compositions on bacterial,bacteriophage, viral, fungal and nematode microbes, using a wide varietyof agricultural products (including leafy vegetables, fruiting andflowering vegetables, podded vegetables, bulb and stem vegetables, rootand tuberous vegetables, Rosaceae family fruits, Bramble fruits,berries, Asian fruit, North America fruit, raw meat, raw poultry, rawseafood, fresh seeds, edible sprouts, and nuts, eggs, fresh herbs, driedherbs, spices, animal hides, feathers) and inanimate objects (such asthose encountered in hospitals, food processing plants, residentialbuildings, office buildings, etc.). Tables 3-5 and 6 also show that theinvention's compositions produced at least a 3-log reduction inmicrobial inoculums that contained microbes ranging from 3-log to8.78-log, after treatment for 2 minutes or longer at a temperature of atleast 34° F. In addition, Tables 5 and 8 show 100% reduction in E. colion spinach and strawberry after 180-minute treatment at about 33° F.with several of the invention's compositions. Additional exemplarycompositions of the invention in Table 6 show 100% reduction in microbeswhen treating inanimate objects.

One surprising property of the invention's compositions is that theyretain their antimicrobial activity over a broad range of temperatures,including from the freezing temperature of the composition to thetemperature of autoclaving (about 120° C.). Thus, the invention'scompositions may be used as antimicrobials at temperatures from thefreezing temperature of the composition to 20° C., to 34° C., to 40° C.,to 60° C., to 80° C., to 100° C. and/or to 120° C. For example, Table 6shows that contacting shrimp, that was contaminated with E. coli, for 30minutes with the invention's compositions that had previously beenfrozen into ice cubes (about 29° F.) resulted in a 6.5-log reduction inE. coli from 6.87-log.

Thus, the invention's compositions advantageously retain theirantimicrobial (and other) activities at temperatures equal to or belowroom temperature. “Room temperature” and “ambient temperature” are usedinterchangeably to refer to 21 degrees celcius (° C.), equivalent to 70degrees Fahrneheit (° F.). “Chilling temperature” refers to atemperature less than room temperature, i.e., less than 21° C.(equivalent to less than 70° F.), including, but are not limited to,temperatures from 30° F. to 69° F., from 34° F. to 45° F., from 34° F.to 69° F., from 40° F. to 42° F., 40° F. to 69° F. and from 50° F. to69° F. Temperatures greater than room temperature include, but are notlimited to, temperatures from 71° F. to 220° F., from 100° F. to 220° F.and from 150° F. to 220° F. Thus, the invention's compositions areparticularly useful at 30° F. to 40° F., preferably at 34° F., which arethe temperatures at which many food products are preferably maintainedprior to human consumption, such as in the field, during transport fromthe field, and before, during and/or after processing for humanconsumption. For agricultural products that may be ingested raw, inseveral embodiments, compositions brought in contact with these productsare preferably formulated to contain one or more GRAS listed component.

Thus, the invention's compositions may be applied to fresh produce andornamental plants at refrigerated temperatures of about 34° F. Inanother embodiment, the invention's compositions may be used whenfrozen, such as ice cubes or crushed ice. The temperature at which thecompositions of the invention freeze will vary depending on the type andconcentration of components in the composition. The invention'scompositions are also useful antimicrobials at room temperature. Forexample in strawberries and tomatoes, a reduction in slime and mold wasobserved over a two-week period of storage at 75° F. following washingwith the invention's compositions.

The invention's compositions are also useful for reducing odorassociated with microbial breakdown of organic material, e.g., food.

In addition, the invention's compositions surprisingly reduceddiscoloration (e.g., pinking and browning) of agricultural products.Thus, in addition to the compositions' preservative effects during atwo-week storage at 40° F. on varieties of lettuce and cabbage, areduction in pinking and browning was also observed when applyinganti-discoloration effective amounts of the invention's compositions. An“anti-discoloration effective amount” of a compound refers to an amountof the compound that reduces the level of discoloration compared to thelevel in the absence of the compound.

“Discoloration” refers to a change in color and/or hue. For example,discoloration of agricultural products includes pinking and browningthat are generally observed after cutting produce and/or during storage.“Pinking” refers to a de novo development and/or increase in pink, redand/or rust color and/or hue such as that observed after storage at thecut edges of lettuce, especially iceberg lettuce, cabbage, celery, andBok Choy. “Browning” refers to de novo development and/or increase inbrown, grey and/or green color and/or hue such as that observed afterstorage at the cut edges of lettuce, cabbage, celery, Bok Choy,potatoes, parsnips, avocado, apples, strawberries, spinach, as well asafter storage of meat (such as hamburger), poultry and seafood.

The level of discoloration may be subjectively determined using knownmethods (Schwank, U.S. Pat. No. 5,087,467) as well as using blind testsas previously described for other attributes such as crispness, color,and appearance of freshness (Estrada, U.S. Pat. No. 5,599,571). Forexample, at 3, 6 and 14 days after treatment with the invention'scompositions, the agricultural produce is rated as having excellent,good, fair or poor color in a blind test by four individuals, and iscompared with produce that is treated under the same conditions with theexception of omission of the invention's composition.

The invention's compositions may be applied by routine methods forapplication of antimicrobial solutions such as immersing, dipping,spraying, fogging, aerosoling, washing and scrubbing. “Fogging” refersto production of liquid droplets of 0.2 μm, or smaller, diameter, e.g.,by applying differential pressure to the liquid. “Aerosoling” refers toproduction of liquid droplets of greater than 0.2 μm diameter, e.g., byapplying high pressure to the liquid.

For large scale processing of produce (e.g., fruits and vegetables), asuitable volume of a working solution of the invention's compositionsmay be contained in a tank or flume, having agitation. The produce isdeposited into this vessel for a period sufficient to reduce microbialcontamination. The treated produce is then removed and transported downthe line for further processing. Final rinsing is not necessary as theamount of residue is very low or non-toxic (Table 9), thus processingmay include a step that leaves only a very thin, adherent film on theproduct's surfaces. This film furnishes an extended period ofantimicrobial activity without adversely altering the color, textureand/or odor of the product surfaces during an extended shelf life.

In addition to the tank or flume, other methods are available by whichthe invention's compositions and their concentrates may be brought tocontact the target microorganisms on any surface, including, withoutlimitation, closed pipes, low-pressure spray, high-pressure spray, fog,vaporization, sonic energy, aerosols, wiping, immersion, scrubbing,swabbing, mopping, brushing, and gels. Contacting the invention'scompositions with target microorganisms on any surface may beaccomplished at atmospheric pressure and/or under a vacuum, i.e., at apressure less than atmospheric pressure.

The invention's compositions may also be used during the processing ofmeat and poultry as previously described (Andrews et al., U.S. Pat. No.5,490,992; Shane et al., U.S. Patent Application US 2002/0134317) andfurther described herein. For example, live animals or poultry arecompletely contacted with one or more of the invention's dilutecompositions using best practices method. The treatment time isdetermined by whether subsequent processing is desired with a wet animalor whether it should appear to be dry. Without rinsing after thisapplication, the invention's composition exhibits residual activity andsubstantially reduces the total microbial load. After the slaughterstep, an animal carcass may be contacted with a more concentratedcomposition of the invention, and most preferably one that contains asurfactant, whereas one or more of the invention's dilute compositionsmay be used in subsequent steps according to best practices. The cuttingand trimming workstations ideally should be cleaned and completelycontacted with one or more of the invention's concentrated compositionsbetween each carcass. In the case of poultry, where cross contaminationis highly possible due to common wash and chiller tanks, one or more ofthe invention's dilute compositions may be used in these tanks. The lastdrainage step may be performed without the need for rinsing and thecarcasses can be packaged normally. The antimicrobial composition isstill effective at the low chilling temperatures and very low amounts.

Seeds and grains used for raw sprouts and/or other ingestion purposesare normally difficult to treat without harm and deterioration. However,contacting these products with one or more of the invention's dilutecompositions and drying without the need for rinsing can substantiallyimprove the product quality and safety.

Additional exemplary uses of the invention's methods are describedherein in the Examples and Table 7 for treating fruits and vegetables,raw eggs, poultry with and without feathers, poultry feathers, meat,hides, seeds and nuts, seafood, dried goods, inanimate objects, skin,reverse osmosis membranes, etc.

After contact with the invention's composition, the treated products maybe rinsed. Alternatively, the treated products may be allowed to dry(e.g., at ambient or chilling temperatures) or dried by spinning,heating, toweling, etc., without rinsing. One advantage of theinvention's compositions is that, after drying, they form a very thinfilm on the surface of the treated products. This film is edible becauseof its low toxicity to higher life forms, while continuing to exhibitantimicrobial activity, thus eliminating the need for rinsing off theinvention's composition from the treated agricultural (or other)products.

The invention's compositions show antibacterial activity against a widespectrum of microbes, including bacterial, viral, fungal and nematodemicrobes. For example, data herein (Tables 3-6 and 8) show theantimicrobial activity of the invention's exemplary compositions on E.Coli (ATCC 25922) and the highly pathogenic E. coli O157:H7 strain (ATCC43888) and Salmonella enterica (ATCC 10708).

The invention's compositions also show antibacterial activity againstGram-positive bacteria. Data herein (Table 6) show the antimicrobialactivity of the invention's exemplary compositions on Staphylococcusaureus (ATCC 25923) and to a lesser extent on Lactobacillus sp. (ATCC55326).

In addition, the invention's compositions show antifungal activity. Dataherein (Table 6) show the antimicrobial activity of the invention'sexemplary compositions on the fungi Aspergillus flavus (ATCC 15517) andSeptoria lycopersici (ATCC Q99324) and toxic mold Stachybotrys chartarum(ATCC 9182).

The invention's compositions also show antiviral activity. Data herein(Table 6) show the antimicrobial activity of the invention's exemplarycompositions on rhinovirus (ATCC vr1110).

Moreover, the invention's compositions show antibacteriophage activity.Data herein (Table 6) show the antimicrobial activity of the invention'sexemplary compositions on bacteriophage T4 (ATCC 35060-B4).

Furthermore, the invention's compositions show antinematode activity.Data herein (Table 6) show the antimicrobial activity of the invention'sexemplary compositions on Aphelenchoides fragariae (ATCC 12974), whichis a nematode that destroys plant crops, necessitating burning of cropsto eradicate the nematode.

One of the surprising properties of the invention's compositions is thatthey show a differential antimicrobial activity on pathogenic andnon-pathogenic microbes. For example, a differential antimicrobialeffect was observed between Gram-positive pathogenic and non-pathogenicbacteria. In particular, Table 6 shows the surprising result thatexemplary composition “x4” of the invention caused greater than a 4-logreduction in a 4.21-log inoculum of Staphylococcus aureus (ATCC 25923)while the exemplary composition “x3” caused less than 2-log reduction ina 8.78-log inoculum of Lactobacillus sp (ATCC 55326).

Also, a differential antimicrobial activity was observed betweenpathogenic Gram-negative bacteria and non-pathogenic Gram-positivebacteria. In particular, Table 6 shows the surprising result thatexemplary compositions x1, x2, x4, x5, x7, x9, x10, x11, x12, x13 andx15 of the invention caused greater than a 6-log reduction in about6.87-log inoculum of E. coli (ATCC 25922 or ATCC 43888) while theexemplary composition “x3” caused less than 2-log reduction in a8.78-log inoculum of Lactobacillus sp (ATCC 55326). Similarly, Table 6shows the surprising result that exemplary compositions x4, x7, x10 andx11 of the invention caused greater than 5-log reduction in a 5.45-loginoculum of S. enterica (ATCC 25922) while the exemplary composition x3caused less than 2-log reduction in a 8.78-log inoculum of Lactobacillussp (ATCC 55326).

Not only was the differential antimicrobial activity observed betweenpathogenic and non-pathogenic bacteria, but it was also observed betweenpathogenic bacteria and non-pathogenic bacteriophage. For example, Table6 shows the surprising result that exemplary compositions “x1” of theinvention caused greater than a 6.8-log reduction in a 6.88-log inoculumof E. coli (ATCC 25922) while the same exemplary composition “x1” causedless than 1-log reduction in a 8.78-log inoculum of bacteriophage T4(ATCC 35060-B4).

Additional differential antimicrobial activity was observed betweenpathogenic and non-pathogenic viruses. For example, Table 6 shows thesurprising result that exemplary compositions “x13” of the inventioncaused a 2.4-log 100% 2.4-log (i.e., 100%) reduction in a 2.4-loginoculum of pathogenic Rhinovirus (ATCC vr1110) while composition “x1”caused less than 1-log reduction in a 8.78-log inoculum ofnon-pathogenic bacteriophage T4 (ATCC 35060-B4).

The invention's compositions are useful as antimicrobials inapplications to a varievariety of surfaces, including agriculturalproducts and inanimate objects.

In one embodiment the agricultural product is a leafy vegetable, asexemplified by, but not limited to, Amaranth (Amaranthus cruentus), Beetgreens (Beta vulgaris subsp. vulgaris), Bitterleaf (Vernonia calvoana),Bok choy (Brassica rapa Chinensis group), Brussels sprout (Brassicaoleracea Gemmifera group), Cabbage (Brassica oleracea Capitata group),Ceylon spinach (Basella alba), Chicory (Cichorium intybus),Chrysanthemum leaves (Chrysanthemum coronarium), Corn salad(Valerianella locusta), Cress (Lepidium sativum), Dandelion (Taraxacumofficinale), Lettuce (Lactuca sativa), Mizuna greens (Brassica rapaNipposinica group), Mustard (Sinapis alba), Napa/Chinese Cabbage(Brassica rapa Pekinensis group), New Zealand Spinach (Tetragoniatetragonioides), Orache (Atriplex hortensis), Pea sprouts/leaves (Pisumsativum), Sea beet (Beta vulgaris subsp. maritima), Seakale (Crambemaritima), Soko (Celosia argentea), Spinach (Spinacia oleracea), Swisschard (Beta vulgaris subsp. cicla var. flavescens), Turnip greens(Brassica rapa Rapifera group), Watercress (Nasturtium officinale),Water spinach (Ipomoea aquatica), and Yau choy (Brassica napus).

In another embodiment, the agricultural product is a fruiting and/orflowering vegetable, as exemplified by, but not limited to, Armeniancucumber (Cucumis melo Flexuosus group), Eggplant or Aubergine (Solanummelongena), Avocado (Persea americana), Bell pepper (Capsicum annuum),Bitter melon (Momordica charantia), Cayenne pepper (Capsicumfrutescens), Chayote (Sechium edule), Chili pepper (Capsicum annuumLongum group), Cucumber (Cucumis sativus), Globe Artichoke (Cynarascolymus), Perennial cucumber (Coccinia grandis), Pumpkin (Cucurbitamaxima, Cucurbita pepo), Pattypan squash, Squash (aka marrow) (Cucurbitapepo), Corn (Zea mays), Sweet pepper (Capsicum annuum Grossum group),Tomato (Solanum lycopersicum), Tomatillo (Physalis philadelphica),Winter melon (Benincasa hispida), and Zucchini or Courgette (Cucurbitapepo).

In a further embodiment, the agricultural product is a podded vegetable,as exemplified by, but not limited to, American groundnut (Apiosamericana), Azuki bean (Vigna angularis), Black-eyed pea (Vignaunguiculata subsp. unguiculata), Chickpea (Cicer arietinum), Fava bean(Vicia faba), Indian pea (Lathyrus sativus), Lentil (Lens culinaris),Mung bean sprouts (Vigna radiata), Okra (Abelmoschus esculentus), Pea(Pisum sativum), Peanut (Arachis hypogaea), Pigeon pea (Cajanus cajan),Rice bean (Vigna umbellatta), Soybean (Glycine max), Winged bean(Psophocarpus tetragonolobus), and Yardlong bean (Vigna unguiculatasubsp. sesquipedalis).

In yet another embodiment, the agricultural product is a bulb vegetableand/or stem vegetable, as exemplified by, but not limited to, Asparagus(Asparagus officinalis), Celery (Apium graveolens), Elephant Garlic(Allium ampeloprasum var. ampeloprasum), Florence fennel (Foeniculumvulgare var. dulce), Garlic (Allium sativum), Kohlrabi (Brassicaoleracea Gongylodes group), Leek (Allium porrum), Onion (Allium cepa),Prussian asparagus (Ornithogalum pyrenaicum), Shallot (Allium cepaAggregatum group), Welsh onion (Allium fistulosum), and Wild leek(Allium tricoccum).

In a further embodiment, the agricultural product is a root vegetableand/or tuberous vegetable, as exemplified by, but not limited to, Bambooshoot, Beetroot (Beta vulgaris subsp. vulgaris), Black cumin (Buniumpersicum), Broadleaf arrowhead (Sagittaria latifolia), Carrot (Daucuscarota), Cassava (Manihot esculenta), Chinese artichoke (Stachysaffinis), Daikon (Raphanus sativus Longipinnatus group), Earthnut pea(Lathyrus tuberosus), Ginger (Zingiber officinale), Hamburg parsley(Petroselinum crispum var. tuberosum), Jerusalem artichoke (Helianthustuberosus), Jicama (Pachyrhizus erosus), Parsnip (Pastinaca sativa),Potato (Solanum tuberosum), Prairie turnip (Psoralea esculenta), Radish(Raphanus sativus), Rutabaga (Brassica napus Napobrassica group), SweetPotato (Kumara), Taro (Colocasia esculenta), Turnip (Brassica rapaRapifera group), Wasabi (Wasabia japonica), Water chestnut (Eleocharisdulcis), and Yam (Dioscorea spp.).

In another embodiment, the agricultural product is a sea vegetable, asexemplified by, but not limited to, Dabberlocks or badderlocks (Alariaesculenta), Dulse (Palmaria palmata), Hijiki (Hizikia fusiformis), Kombu(Laminaria japonica), Mozuku (Cladosiphon okamuranus), Sea grape(Caulerpa spp.), and Sea lettuce (Ulva lactuca).

In other embodiments, the agricultural product is a member of theRosaceae family, as exemplified by, but not limited to, Apple andcrabapple (Malus), Hawthorn (Crataegus and Rhaphiolepis), Loquat(Eryobotrya japonica), Pear, European and Asian species (Pyrus), Quince(Cydonia oblonga and Chaenomeles), Apricot (Prunus armeniaca orArmeniaca vulgaris), Cherry (Prunus avium), Peach (Persica, vulgaris),Plum (Prunus salicina), and Strawberry (Fragaria ananassa)

In another embodiment, the agricultural product is a bramble fruit, asexemplified by, but not limited to, Blackberry (genus Rubus) andRaspberry (genus Rubus), or is a berry, as exemplified by, but notlimited to, Bilberry or whortleberry (Vaccinium spp.), Blueberry(Vaccinium spp.), Cranberry (Vaccinium spp.), Huckleberry (Vacciniumspp.), Barberry (Berberis; Berberidaceae), Currant (Ribes spp.;Grossulariaceae) including red, black, and white types, Elderberry(Sambucus; Caprifoliaceae), Gooseberry (Ribes spp.; Grossulariaceae),Hackberry (Celtis spp.; Cannabaceae), Honeysuckle, (Lonicera spp.;Caprifoliaceae), and Mulberry (Morus spp.; Moraceae).

In a further embodiment, the agricultural product is an Asian fruit, asexemplified by, but not limited to, Coconut (Cocos spp.; Arecaceae),Kiwifruit or Chinese gooseberry (Actinidia spp.; Actinidiaceae),Persimmon (aka Sharon Fruit) (Diospyros kaki; Ebenaceae), Rhubarb (Rheumrhaponticum; Polygonaceae), and Pineapple (Ananas comosus).

In another embodiment, the agricultural product is a North Americanfruit, as exemplified by, but not limited to, American grape, (Vitislabrusca; Vitaceae), American persimmon (Diospyros virginiana;Ebenaceae), Beach Plum (Prunus maritima; Rosaceae), Blueberry(Vaccinium, sect. Cyanococcus; Ericaceae), Persimmon (Diospyrosvirginiana; Ebenaceae), and Saw Palmetto (Serenoa repens; Ericaceae).

In yet another embodiment, the agricultural product is a citrus fruit,as exemplified by, but not limited to, Blood Orange, Citron (Citrusmedica), Clementine (Citrus reticulata var. Clementine), Grapefruit(Citrus paradisi), Kumquat (Fortunella), Lemon (Citrus limon), Key Lime(Citrus aurantifolia), Mandarin (Citrus reticulata), Orange, (Citrussinensis), Pomelo (also known as the shaddock) (Citrus maxima), andSweet Lemon (Citrus limetta).

In further embodiments, the agricultural product is meat, such as rawand/or cooked beef, pork, and lamb, exemplified by beef steak, beefhamburger, pork chop, pork sausage, lamb shank, and bacon.

In other embodiments, the agricultural product is poultry, such as rawand/or cooked turkey, chicken, and duck, including whole poultry andpoultry parts, e.g., breast, thigh, drumstick, and poultry productse.g., poultry eggs, and egg shells.

In yet other embodiments, the agricultural product is seafood such asraw (e.g., sushi) and/or cooked fish (e.g., salmon, bass, tuna),including whole fish and fish parts, e.g., steak, fillet, as well asshellfish (e.g., mussels, clams, oysters, and shrimp).

In further embodiments, the agricultural product is a fresh or cookedseed or nut, as exemplified by, but not limited to, almonds (Prunuscommunis), walnuts (Juglans nigra), sunflower (Helianthus, annuus), andpumpkin (Cucurbita pepo).

Other embodiments of agricultural products include fresh and/or driedherb such as parsley (Carum petroselinum), cumin (Cuminum cyminum),sweet basil (Ocimum basilicum), rosemary (Rosmarinus officinalis), andmint (Mentha spp).

Further embodiments of agricultural product include animal hide (e.g.,cow hide and pig hide), and poultry feathers (e.g., chicken feathers,turkey feathers and goose feathers).

Other surfaces that may be treated with the invention's compositionsinclude skin (e.g., human skin, cow udders, cow teats, etc.) andinanimate objects (e.g., door handles, door knobs, door frames, tablesurfaces, stair railings).

EXPERIMENTAL

The following examples serve to illustrate certain preferred embodimentsand aspects of the present invention and are not to be construed aslimiting the scope thereof

EXAMPLE 1 Synergistic Antimicrobial Activity

This example provides the results for antimicrobial activity ofcomponents of exemplary invention's compositions when the componentswere used alone (Tables 3A and 3B), or as a combination of twocomponents (Tables 3A and 4), or of at least three components (Tables 3Aand 5).

Produce was rinsed with sterile water before refrigerating. Produce wasdip-inoculated and let stand for 24 hrs prior to component testing.25-gram samples were pureed with 475 grams of sterile water (20 folddilution). Pureed samples were spread-plated on pre-made Mac Conkey withmug agar plates #221172 from Becton, Dickinson Biosciences and incubatedat about 44° F. for 24-30 hours. Colonies were counted using colonycounter Bantex 920A with UV light 366 nM. The results are shown in Table3A.

TABLE 3A Antimicrobial activity (log reduction) of components usedsingly, in combination of two components, in combination of threecomponents, and in combination of four components. E. Coli (ATCC 29522)inoculum 5.45 log Strawberry 25 grams Spinach leaf 25 gramsComponent^((a)) 2 min.^((b)) 30 min.^((b)) 180 min.^((b)) 2 min.^((b))30 min.^((b)) 180 min.^((b)) Single Components Propylene Glycol (0.03wgt. 1.83 2.24 2.55 1.71 2.1 2.4 %) Phosphoric Acid (0.3 wgt. 2.15 2.172.62 2.00 2.1 2.55 %) Hydrogen Peroxide (0.2 wgt. 2.0 2.66 2.99 1.8 2.442.77 %) Sodium Lauryl Sulfate 1.2 1.5 1.89 1.0 1.6 1.78 (0.0005 wgt. %)Potable Water alone 0.91 0.98 1.05 0.78 0.82 0.88 (33° F., 100 wgt. %)Two Components Propylene Glycol (0.03 wgt. 2.99 4.01 5.45 2.78 3.89 5.45%) and Phosphoric Acid (0.3 wgt. %) Three Components Propylene Glycol(0.03 wgt. 3.34 4.45 5.45 3.12 4.12 5.45 %) and Phosphoric Acid (0.3wgt. %) and Hydrogen Peroxide (0.2 wgt. %) Four Components PropyleneGlycol (0.03 wgt. 4.12 5.11 5.45 3.98 4.99 5.45 %) and Phosphoric Acid(0.3 wgt. %) and Hydrogen Peroxide (0.2 wgt. %) and Sodium LaurylSulfate (0.0005 wgt. %)^((c)) ^((a))Component was dissolved in potablewater to make a total of 100 g. The component and sterile water werechilled to 34° F. ^((b))Fruit and vegetable were chilled to 40° F. andwashed in the solution containing the listed component for the periodindicated. Testing was done at ambient temperature of about 68° F.^((c))The solution containing Propylene Glycol 0.03 wgt. %,ortho-Phosphoric Acid 0.3 wgt. %, Hydrogen Peroxide 0.2 wgt. % andSodium Lauryl Sulfate 0.0005 wgt. % is an exemplary composition ofsolution D of Table 1. It was made by using a concentrated solution ofPropylene Glycol 3 wgt. %, ortho-Phosphoric Acid 30 wgt. %, HydrogenPeroxide 20 wgt. % and Sodium Lauryl Sulfate 0.05 wgt. % diluted 100 to1.

TABLE 3B Antimicrobial activity (log reduction) of single components E.Coli (ATCC 43888) inoculum 6.87-log Strawberry 25 grams Spinach leaf 25grams Component^((a)) 2 min.^((b)) 30 min.^((b)) 180 min.^((b)) 2min.^((b)) 30 min.^((b)) 180 min.^((b)) Propylene Glycol (2 wgt. %) 2.232.69 4.47 2.44 2.54 3.98 Acetic Acid (1 wgt. %) 2.52 2.73 4.45 2.5 2.614.01 Citric Acid (1 wgt. %) 2.25 2.68 4.43 2.11 2.34 4.09 PhosphoricAcid (1 wgt. %) 2.58 2.87 4.22 2.5 2.71 4.13 Hypochlorous Acid (0.01wgt. %) 2.45 3.45 5.69 2.32 3.4 5.01 Bromine (0.08 wgt. %) 2.32 3.324.89 2.11 3.28 4.44 Hydrogen Peroxide (2 wgt. %) 2.11 2.69 4.65 2.01 2.63.89 Ozone (0.001 wgt. %) 2.89 3.12 4.12 2.34 2.58 3.63 Sodium LaurylSulfate 1.89 2.69 3.56 1.77 2.29 3.26 (0.5 wgt. %) (CAS 009004-82-4)Octyl Phenol Ethoxylate 1.05 2.78 3.9 1.15 2.47 3.11 (0.5 wgt. %)Potable Water alone 0.98 1.04 1.2 0.68 1 1.1 ^((a))Component wasdissolved in potable water to make a total of 100 g and tested at about33° F. ^((b))Fruit and vegetable were washed in the solution containingthe listed component for the period indicated.

TABLE 4 Antimicrobial activity (log reduction) of a combination of twocomponents E. Coli (ATCC 43888) inoculum 6.87-log Strawberry 25 gramsSpinach leaf 25 grams Component^((a)) 2 min.^((b)) 30 min.^((b)) 180min.^((b)) 2 min.^((b)) 30 min.^((b)) 180 min.^((b)) Hypochlorous Acid0.01 wgt. % 2.68 3.74 4.98 2.48 3.54 4.78 & Acetic Acid 1 wgt. %Hypochlorous Acid 0.01 wgt. 2.55 3.51 4.66 2.25 3.3 4.48 % & Citric Acid1 wgt. % Hypochlorous Acid 0.01 wgt. 2.81 3.94 5.13 2.61 3.74 4.9 % &Phosphoric Acid 1 wgt. % Bromine 0.08 wgt. % & 2.71 3.39 4.91 2.51 3.14.7 Acetic Acid 1 wgt. % Bromine 0.08 wgt. % & 2.44 3.64 4.71 2.52 3.544.71 Citric Acid 1 wgt. % Bromine 0.08 wgt. % & 2.82 3.93 4.74 2.42 3.634.88 Phosphoric Acid 1 wgt. % Hydrogen Peroxide 2 wgt. 2.51 3.56 4.892.43 3.5 4.77 % & Acetic Acid 1 wgt. % Hydrogen Peroxide 2 wgt. 2.543.57 4.85 2.4 3.68 4.85 % & Citric Acid 1 wgt. % Hydrogen Peroxide 2wgt. 2.72 3.81 4.99 2.69 3.71 4.99 % & Phosphoric Acid 1 wgt. % Ozone0.001 wgt. % & 2.49 3.79 4.96 2.65 3.5 4.9 Acetic Acid 1 wgt. % Ozone0.001 wgt. % & 2.48 3.77 4.96 2.48 3.57 4.7 Citric Acid 1 wgt. % Ozone0.001 wgt. % & 2.47 3.78 4.97 2.5 3.7 4.78 Phosphoric Acid 1 wgt. %Hypochlorous Acid 0.01 wgt. 2.41 3.31 4 2.61 3.41 4.1 % & Sodium LaurylSulfate 0.5 wgt. % Hypochlorous Acid 0.01 wgt. 2.56 3.41 4.11 2.4 3.21 4% & Octyl Phenol Ethoxylate 0.5 wgt. % Bromine 0.08 wgt. % & 2.31 2.983.98 2.3 2.88 3.99 Sodium Lauryl Sulfate 0.5 wgt. % Bromine 0.08 wgt. %& 2.45 2.99 4 2.4 2.9 3.78 Octyl Phenol Ethoxylate 0.5 wgt. % HydrogenPeroxide 2 wgt. 2.34 2.89 3.57 2.35 2.88 3.55 % & Sodium Lauryl Sulfate0.5 wgt. % Hydrogen Peroxide 2 wgt. 2.46 2.93 3.91 2.33 2.78 3.81 % &Octyl Phenol Ethoxylate 0.5 wgt. % Ozone 0.001 wgt. % & 2.56 3.51 3.982.56 3.31 3.78 Sodium Lauryl Sulfate 0.5 wgt. % Ozone 0.001 wgt. % &2.54 3.41 3.9 2.45 3.22 3.39 Octyl Phenol Ethoxylate 0.5 wgt. %^((a))Component was dissolved in potable water to make a total of 100 gand tested at about 33° F. ^((b))Fruit and vegetable were washed in thesolution containing the listed component for the period indicated.

TABLE 5 Antimicrobial activity (log reduction) of a combination of atleast three components. E. Coli (ATCC 43888) inoculum 6.57-logStrawberry 25 grams Spinach leaf 25 grams Component^((a)) 2 min.^((b))30 min.^((b)) 180 min.^((b)) 2 min.^((b)) 30 min.^((b)) 180 min.^((b))(a1) 4 5.1 6.57 4.5 5.2 6.57 Hypochlorous Acid 0.01 wgt. % & PhosphoricAcid 1 wgt. % & Propylene Glycol 2 wgt. % (a2) 3.9 4.48 6.49 4 4.58 6.49Bromine 0.08 wgt. % & Phosphoric Acid 1 wgt. % & Propylene Glycol 2 wgt.% (a3) 3.98 4.22 6.5 3.99 4.32 6.5 Hydrogen Peroxide 2 wgt. % &Phosphoric Acid 1 wgt. % & Propylene Glycol 2 wgt. % (a4) 3.99 4.77 6.493.99 4.76 6.48 Ozone 0.001 wgt. % & Phosphoric Acid 1 wgt. % & PropyleneGlycol 2 wgt. % (a5) 4.12 5.44 6.57 4.22 5.3 6.57 Hypochlorous Acid 0.01wgt. % & Bromine 0.05 wgt. % & Phosphoric Acid 1 wgt. % & PropyleneGlycol 2 wgt. % ^((a))Components were dissolved in potable water to makea total of 100 g and tested at about 33° F. ^((b))Fruit and vegetablewere washed in the solution containing the listed components for theperiod indicated.

In the above Tables 3-5, the antimicrobial activity is reported as a logreduction which was determined by calculating the difference between thelog₁₀ of the initial inoculum count and the log₁₀ of the inoculum'scount after exposure to the listed components for about 2 minutes, about30 minutes, or about 180 minutes, at about 33° F.

To determine bacterial kill-rate or log₁₀ reduction, a 0.1 ml aliquot ofa bacterial culture suspension incubated for about 24 hours in trypticsoy broth having an initial inoculum count of between about 10⁶ to about10⁸ cells/ml was added to a test sample of either fruits (strawberry 25grams) or vegetables (spinach 25 grams) at about 33° F. In this example,culture suspensions were prepared from E. coli (ATCC 43888). After abouttwo minutes, thirty minutes or one hundred eighty minutes of thistreatment, using the USDA recommended procedure, the inoculated materialwas placed into a 225 ml solution of EC Medium, with mug, in a stomacherbag and thoroughly mixed for a minimum of two minutes and incubated at35° C. for twenty four hours. Next, the cultures were diluted 10-fold inButterfield's Phosphate Diluent and 0.1 ml of the dilutions wereinoculated using the spread plate technique onto MacConkey Sorbitol Agar(MSA). Then the plates were incubated for twenty four hours at 42° C.Plates were then removed and examined for the number of colonies.

The data demonstrate synergy of the invention's components. For example,Table 3B shows that 180 minute treatment of spinach with each ofhypochlorous acid 0.01 wgt. %, phosphoric acid 1 wgt. % and propyleneglycol 2 wgt. % alone in water caused a 5.01-log, 4.13-log, and 3.98-logreduction, respectively, in a 6.87-log E. coli inoculum. Table 4 showssimilarly low antimicrobial activity by a combination of two componentsof hypochlorous acid 0.01 wgt. % and phosphoric acid 1 wgt. % whichresulted in a 4.9-log reduction in a 6.87-log E. coli inoculum. Incontrast, Table 5 shows that 180 minute treatment of spinach with thecombination “a1” that contained hypochlorous acid 0.01 wgt. %,phosphoric acid 1 wgt. %, and propylene glycol 2 wgt. % resulted in a100% reduction in a 6.57-log E. coli inoculum.

Similarly, Table 3B shows that 180 minute treatment of spinach with eachof bromine 0.08 wgt. %, phosphoric acid 1 wgt. % and propylene glycol 2wgt. % alone caused a 4.44-log, 4.13-log, and 3.98-log reduction,respectively, in a 6.87-log E. coli inoculum. Table 4 shows similarlylow antimicrobial activity by a combination of two components of bromine0.08 wgt. % and phosphoric acid 1 wgt. % which resulted in a 4.88-logreduction in a 6.87-log E. coli inoculum. In contrast, Table 5 showsthat 180 minute treatment of spinach with the combination “a2” thatcontained bromine 0.08 wgt. %, phosphoric acid 1 wgt. % and propyleneglycol 2 wgt. % resulted in a 6.49-log reduction in a 6.57-log E. coliinoculum.

In yet another example, Table 3B shows that 180 minute treatment ofspinach with each of hypochlorous acid 0.01 wgt. %, bromine 0.05 wgt. %,phosphoric acid 1 wgt. %, and propylene glycol 2 wgt. % alone caused a5.01-log, 4.44-log, 4.13-log and 3.98-log reduction, respectively, in a6.87-log E. coli inoculum. Table 4 shows similarly low antimicrobialactivity by a combination of two components of bromine 0.08 wgt. % andphosphoric acid 1 wgt. % which resulted in a 4.88-log reduction in a6.87-log E. coli inoculum. In contrast, Table 5 shows that 180 minutetreatment of spinach with the combination “a5” that containedhypochlorous acid 0.01 wgt. %, bromine 0.05 wgt. %, phosphoric acid 1wgt. %, and propylene glycol 2 wgt. % resulted in a 100% reduction in a6.57-log E. coli inoculum.

EXAMPLE 2 Antimicrobial Activity Using Agricultural Products andInanimate Objects

This example shows the results of treating a variety of agriculturalproducts, skin, and inanimate objects with the invention's exemplarycompositions “x1” to “x15” that are described above in Table 2.

TABLE 6 Antimicrobial Activity Of Exemplary Compositions Of TheInvention Reference Treatment Log Number of Method^((a)) Log ReductionExemplary (see Table 7 Microbe in Material Composition for details)Microbe Tested^((b)) Inoculum Microbe^((c)) Notes Botanical Leafyvegetables Bok choy x1 t1 b2 6.88 >6.7 (Brassica rapa (E. coli generic)Chinensis group) x3 t3 b4 8.78 <2.3 Composition x3 (Lactobacillus sp.)diluted 100 to 1 Cabbage x1 t1 b2 6.88 >6.8 (Brassica (E. coli generic)oleracea Capitata group) x3 t3 b4 8.78 <1 Composition x3 (Lactobacillussp.) diluted 100 to 1 x15 t2 b1 6.87 >6.7 Composition (E. coli) x15diluted 10 to 1 Dandelion x1 t1 b2 6.88 >6.8 (Taraxacum (E. coligeneric) officinale) x3 t3 b4 8.78 <2 Composition x3 (Lactobacillus sp.)diluted 100 to 1 Lettuce x1 t1 b2 6.88 >6.8 (Lactuca sativa) (E. coligeneric) x3 t3 b4 8.78 <3 Composition x3 (Lactobacillus sp.) diluted 100to 1 x4 t2 b3 4.21 >4 (Staphylococcus aureus) x4 t2 b5 5.45 >5(Salmonella enterica) Mustard x1 t1 b2 6.88 >6.8 (Sinapis alba) (E. coligeneric) x3 t3 b4 8.78 <1 Composition x3 (Lactobacillus sp.) diluted 100to 1 Napa/Chinese x1 t1 b2 6.88 >6.6 Cabbage (E. coli generic) (Brassicarapa Pekinensis group) x3 t3 b4 8.78 <1 Composition x3 (Lactobacillussp.) diluted 100 to 1 Spinach x1, t1 b2 6.88 >6.7 (Spinacia (E. coligeneric) oleracea) x3 t3 b4 8.78 <2 Composition x3 (Lactobacillus sp.)diluted 100 to 1 x11 t3 b1 6.87 >6.5 Composition (E. coli) x11 diluted200 to 1 x7 t2 b5 5.45 >5 (Salmonella enterica) Swiss chard x1 t1 b26.88 >6.8 (Beta vulgaris (E. coli generic) subsp. cicla var. flavescens)x3 t3 b4 8.78 <1 Composition x3 (Lactobacillus sp.) diluted 100 to 1 x9t1 b2 6.88 >6.6 (E. coli generic) x1 t1 p1 3.1 <1 (bacteriophage T4)Botanical Fruiting and flowering vegetables Avocado x1 t3 b1 6.87 >6.7(Persea (E. coli) americana) x2 t1 b1 6.87 >6 (E. coli) Bell pepper x4t3 b1 6.87 >6.8 Composition x4 (Capsicum (E. coli) diluted 100 to 1annuum) x9 t1 b1 6.87 >6.7 (E. coli) Globe x5 t3 b1 6.87 >6.6 Artichoke(E. coli) (Cynara scolymus) x7 t2 b1 6.87 >6.8 (E. coli) Tomato x1 t1 b26.88 >6.8 (Solanum (E. coli generic) lycopersicum) x2 t2 b5 5.45 <2(Salmonella enterica) x11 t3 m1 5.5 >4 Composition (Stachybotrys x11diluted 100 chartarum) to 1 x1 t2 b2 6.88 >6.6 (E. coli generic) x13 t3m1 5.5 >5 Composition (Stachybotrys x13 diluted 100 chartarum) to 1Botanical Podded vegetables Mung bean x1 t1 b1 6.87 >6.5 sprouts (Vigna(E. coli) radiata) x4 t3 b1 6.87 >6.1 Composition x4 (E. coli) diluted100 to 1 Botanical Bulb and stem vegetables Asparagus x10 t3 b16.87 >6.5 (Asparagus (E. coli) officinalis) Botanical Root and tuberousvegetables Carrot (Daucus x10 t3 b1 6.87 >6.6 carota) (E. coli)Botanical Rosaceae family Apple and x10 t3 b1 6.87 >6.6 crabapple (E.coli) (Malus) Peach x10 t3 b1 6.87 >6.5 (Persica, (E. coli) vulgaris)Strawberry x1 t3 b1 6.87 >6.7 (Fragaria (E. coli) ananassa) x4 t1 b26.88 >6.6 Composition x4 (E. coli generic) diluted 100 to 1 x11 t2 m15.5 >4 Composition (Stachybotrys x11 diluted 50 chartarum) to 1Botanical Bramble fruits Blackberry, x11 t3 b1 6.87 >6.8 Composition(genus Rubus) (E. coli) x11 diluted 100 to 1 Botanical Berries Cranberryx11 t3 b1 6.87 >6.7 Composition (Vaccinium (E. coli) x11 diluted 100spp.) to 1 Botanical Fruit Asian Pineapple cut x11 t3 b1 6.87 >5Composition into (E. coli) x11 diluted 500 slices(Ananas to 1 comosus)Botanical Fruit North American Blueberry x11 t3 b1 6.87 >6 Composition(Vaccinium, (E. coli) x11 diluted 100 sect. to 1 Cyanococcus; Ericaceae)Meat Raw Beef x1 t8 b1 6.87 >5 Hamburger (E. coli) Pork Chop x11 t9 b16.87 >6 Composition (E. coli) x11 diluted 100 to 1 Bacon x1 t8 b16.87 >6.5 (E. coli) Poultry Raw Turkey breast x11 t6 b1 6.87 >6Composition (E. coli) x11 diluted 100 to 1 Chicken breast x1 t6 b16.87 >6.8 (E. coli) Fish Raw Salmon steak x1 t13 b1 6.87 >6.8 (E. coli)Bass steak x12 t13 b1 6.87 >6.8 (E. coli) tuna processed x12 t13 b16.87 >6 raw (E. coli) Shellfish Clams raw x1 t14 b1 6.87 >6 (E. coli)shrimp raw not x1 t14 b1 6.87 >6.5 frozen (E. coli) Seeds and Nuts fresh(not dried) Almonds x12 t11 m1 5.5 >4 (Prunus (Stachybotrys communis)chartarum) x12 t11 f1 2.3 >1.8 (Aspergillus flavus) Eggs (topicalShells) Chicken fresh x15 t4 b1 6.87 >6 eggs (E. coli) Herbs freshparsley (Carum x1 t1 b1 6.87 >6 petroselinum) (E. coli) Herbs Driedparsley (Carum x13 t15 b1 6.87 >6 Composition petroselinum) (E. coli)x13 diluted 200 to 1 Animal Hides Cow Hide x11 t10 b1 6.87 >6Composition (E. coli) x11 diluted 50 to 1 Animal feathers Chicken x11 t7b5 5.45 >5 feathers (Salmonella enterica) Human skin Male age 46, x10t19 skin test — forearm Male age 78, x10 t19 b3 (Staphylococcus 4.21 —top left hand aureus) Structures Pine wood x5 t18 m1 5.5 >3.5 sanded 6″× 6″ (Stachybotrys chartarum) x9 t18 b1 6.87 >5 (E. coli) x14 t18 b26.88 >5 (E. coli generic) Teflon sheet x5 t18 m1 5.5 >5 6″ × 6″(Stachybotrys chartarum) x9 t18 b1 6.87 >6 (E. coli) x14 t18 b2 6.88 >6(E. coli generic) Ultra high x5 t18 m1 5.5 >5 molecular (Stachybotrysweight chartarum) (UHMW) polyethylene sheet 6″ × 6″ x9 t18 b1 6.87 >6(E. coli) x14 t18 b2 6.88 >6 (E. coli generic) ceramic tile x13 t17 m15.5 5.5 Composition 3″ × 3″ (Stachybotrys x13 diluted 10 chartarum) to 1x13 t17 v1 2.4 2.4 Composition (Rhinovirus) x13 diluted 10 to 1stainless steel x13 t17 m1 5.5 5.5 Composition 6″ × 6″ (Stachybotrys x13diluted 10 chartarum) to 1 x13 t17 v1 2.4 2.4 Composition (Rhinovirus)x13 diluted 10 to 1 cement block x13 t17 m1 5.5 5.5 Composition 6″ × 6″(Stachybotrys x13 diluted 10 chartarum) to 1 x13 t17 v1 2.4 2.4Composition (Rhinovirus) x13 diluted 10 to 1 Glass sheet x13 t17 m1 5.55.5 Composition 6″ × 6″ (Stachybotrys x13 diluted 10 chartarum) to 1 x13t17 v1 2.4 2.4 Composition (Rhinovirus) x13 diluted 10 to 1 Painted(acrylic x13 t18 m1 5.5 >4 Composition paint) wood (Stachybotrys x13diluted 10 6″ × 6″ chartarum) to 1 x13 t18 v1 2.4 >2 Composition(Rhinovirus) x13 diluted 10 to 1 ICE with x1 t16 b1 6.87 >6 shrimp (E.coli) x2 t16 b1 6.87 >6.5 (E. coli) Plant specific Tomato leaf x11 t1 f23.2 >2 Composition (Solanum (Septoria x11 diluted 50 lycopersicum)lycopersici) to 1 x11 t1 n1 2.3 >1.8 Composition (Aphelenchoides x11diluted 50 fragariae) to 1 Poultry x10 t7 b5 5.45 >5 feathers(Salmonella enterica) Reverse x3 t21 b2 6.88 6.88 Composition x3 Osmosis(RO) (E. coli generic) diluted 200 to 1 membrane ^((a))All tests wereperformed according to FDA specifications. ^((b))Microbes: E. coli (ATCC43888), E. coli generic (ATCC 25922), Staphylococcus aureus (ATCC25923), Salmonella enterica (ATCC 10708), Lactobacillus sp. (ATCC55326). Aspergillus flavus (ATCC 15517) is an almond fungus; Septorialycopersici (ATCC Q99324) is a tomato leaf fungus; Stachybotryschartarum (ATCC 9182) is black mold (toxic); bacteriophage T4 (ATCC35060-B4); Aphelenchoides fragariae (ATCC 12974) is a nematode thatdestroys plant crops, necessitating burning of crops to eradicate thenematode; rhinovirus (ATCC vr1110) causes head cold in humans. ^((c))<means less than; > means more than.

TABLE 7 Exemplary Treatment Methods Used To Obtain Some Of The Data InTable 6 Treatment Reference Treatment Method^((a)) t1 Fruits andvegetables may be dipped, sprayed, fogged or aerosoled at temperaturesequal to or above 40 degrees F. as a pretreatment before the finalprocessing wash. Contact time with the invention's composition may be 45minutes or longer. t2 Fruits and vegetables may be dipped, sprayed,fogged or aerosoled at temperatures equal to or below 40 degrees F. as apretreatment before the final processing wash. Contact time with theinvention's composition may be 100 minutes or longer. t3 Fruits andvegetables may be liquid conveyed in composition at temperatures equalto or below 40 degrees F. as a final processing wash. Contact time withthe invention's composition may be 100 minutes or longer. t4 Raw eggsmay be dipped, sprayed, fogged or aerosoled at temperatures equal to orabove 34 degrees F. as a final treatment before packaging. Contact timewith the invention's composition may be 45 minutes or longer. t5 Poultrywith feathers may be dipped, sprayed, fogged or aerosoled attemperatures equal to or above 70 degrees F. as a pretreatment beforede-feathering. Contact time with the invention's composition may be 15minutes or longer. Feathers will have reduced numbers of microbes andcan be dried and ready for processing. for use as fertilizer. t5 Poultrywith feathers may be dipped, sprayed, fogged or aerosoled attemperatures equal to or above 70 degrees F. as a pretreatment beforede-feathering. Contact time with the invention's composition may be 15minutes or longer. Feathers will have reduced numbers of microbes andcan be dried and ready for processing. t6 Poultry that has beende-feathered may be dipped, sprayed, fogged or aerosoled at temperaturesequal to or above 34 degrees F. as a final treatment before packaging.Contact time with the invention's composition may be 45 minutes orlonger. t6 Poultry that has been de-feathered may be dipped, sprayed,fogged or aerosoled at temperatures equal to or above 34 degrees F.Contact time with the invention's composition may be 45 minutes orlonger. t7 Detached Ppoultry feathers may be dipped, sprayed, fogged oraerosoled at temperatures equal to or above 70 degrees F. as a finaltreatment before palletizing.further use. Contact time with theinvention's composition may be 5 minutes or longer. Feathers will havereduced numbers of microbes and residual activity even after can bedried and are readydrying. for processing for use as fertilizer. t7Detached poultry feathers may be dipped, sprayed, fogged or aerosoled attemperatures equal to or above 70 degrees F. as a final treatment beforefurther use. Contact time with the invention's composition may be 5minutes or longer. Feathers will have reduced numbers of microbes andresidual activity even after drying. t8 Meats other than poultry, may bedipped, sprayed, fogged or aerosoled at temperatures equal to or above34 degrees F. as a final wash. Contact time with the invention'scomposition may be 45 minutes or longer. t9 Meats other than poultry,maybe dipped, sprayed, fogged or aerosoled at temperatures equal to orabove 34 degrees F. as a final wash. Contact time with the invention'scomposition may be 45 minutes or longer. The composition is preferablywashedrinsed off. t9 Meats other than poultry, maybe dipped, sprayed,fogged or aerosoled at temperatures equal to or above 34 degrees F. as afinal wash. Contact time with the invention's composition may be 45minutes or longer. The composition is preferably rinsed off. t10 Hides(animal skins) may be dipped, sprayed, fogged or aerosoled attemperatures equal to or above 34 degrees F. as a pretreatment. Contacttime with the invention's composition may be 5 minutes or longer.Animals may be alive or dead for this process. The composition may beleft on to dry before the remainder of the processing steps. t11 Seedsand nuts may be dipped, sprayed, fogged or aerosoled at temperaturesequal to or above 34 degrees F. as a final wash. Contact time with theinvention's composition may be 45 minutes or longer. t12 Seeds and nutsmay be dipped, sprayed, fogged or aerosoled at temperatures equal to orabove 34 degrees F. as a final wash. Contact time with the invention'scomposition may be 15 minutes or longer. The composition is preferablywashedrinsed off. t12 Seeds and nuts may be dipped, sprayed, fogged oraerosoled at temperatures equal to or above 34 degrees F. as a finalwash. Contact time with the invention's composition may be 15 minutes orlonger. The composition is preferably rinsed off. t13 Fish may bedipped, sprayed, fogged or aerosoled at temperatures equal to or above34 degrees F. as a final wash. Contact time with the invention'scomposition may be 15 minutes or longer. t14 Shellfish may be dipped,sprayed, fogged or aerosoled at temperatures equal to or above 34degrees F. as a final wash. Contact time with the invention'scomposition may be 15 minutes or longer. t15 Dried goods may be sprayed,fogged or aerosoled at temperatures equal to or above 34 degrees F. as afinal wash. Contact time with the invention's composition may be Contacttime with the invention's composition may be 115 minutes or longer. t15Dried goods may be sprayed, fogged or aerosoled at temperatures equal toor above 34 degrees F. as a final wash. Contact time with theinvention's composition may be 15 minutes or longer. t16 Ice may beformed with the composition that is to be used as an aid for coldstorage transport and microbial reduction. The shrimp were dipped into asolution containing a bacterial inoculum, and then surrounded for 30minutes at ambient temperature (about 62° F.) by the invention'scompositions that had previously been frozen into ice cubes (about 29°F.). Materials surrounded by the ice could be rinsed off, or leftwithout washing offrinsing to allow formation of a bio-thin, adherentfilm that provides continued antimicrobial activity. t16 Ice may beformed with the composition that is to be used as an aid for coldstorage transport and microbial reduction. The shrimp were dipped into asolution containing a bacterial inoculum, and then surrounded for 30minutes at ambient temperature (about 62° F.) by the invention'scompositions that had previously been frozen into ice cubes (about 29°F.). Materials surrounded by the ice could be rinsed off, or leftwithout rinsing to allow formation of a thin, adherent film thatprovides continued antimicrobial activity. t17 Structures may be dipped,sprayed, fogged, aerosoled, autoclaved or scrubbed in at temperaturesequal to or greater than 200 degrees F. as a final wash. Contact timewith the invention's composition may be 2 minutes or longer. Thecomposition is preferably left on the surface without washing offrinsingto allow formation of a bio-thin, adherent film that provides continuedantimicrobial activity t17 Structures may be dipped, sprayed, fogged,aerosoled, autoclaved or scrubbed at temperatures equal to or greaterthan 200 degrees F. as a final wash. Contact time with the invention'scomposition may be 2 minutes or longer. The composition is preferablyleft on the surface without rinsing to allow formation of a thin,adherent film that provides continued antimicrobial activity t18Structures may be dipped, sprayed, fogged, aerosoled or scrubbed in attemperatures equal to or greater than 34 degrees F. as a final wash.Contact time with the invention's composition may be 10 minutes orlonger. t18 Structures may be dipped, sprayed, fogged, aerosoled orscrubbed at temperatures equal to or greater than 34 degrees F. as afinal wash. Contact time with the invention's composition may be 10minutes or longer. t19 Human skin may be washed at temperatures equal toor above 34 degrees F. as an antimicrobial wash. Contact time with theinvention's composition may be 2 minutes or longer. cContact time of4-12 hours did not result in skin irritation. The moist composition maybe lightly toweled off.for a dry appearance. t19 Human skin may bewashed at temperatures equal to or above 34 degrees F. as anantimicrobial wash. Contact time with the invention's composition may be2 minutes or longer. Contact time of 4-12 hours did not result in skinirritation. The moist composition may be lightly toweled for a dryappearance. t20 Human skin may be washed at temperatures equal to orabove 34 degrees F. as an antimicrobial wash. Contact time with theinvention's composition may be 15 minutes or longer. The composition ispreferably washedrinsed off. t20 Human skin may be washed attemperatures equal to or above 34 degrees F. as an antimicrobial wash.Contact time with the invention's composition may be 15 minutes orlonger. The composition is preferably rinsed off. t21 The invention'scomposition was circulated at temperatures equal to or above 34 degreesF. through thea RO membrane for 1 minute using a high pressure multiplecentrifuge pump. t21 The invention's composition was circulated attemperatures equal to or above 34 degrees F. through a RO membrane for 1minute using a high pressure multiple centrifuge pump. ^((a))Contacttime with the invention's compositions refers to the period of time thecomposition is in contact with a substance before rinsing, drying, ortoweling.

The above Table 6 shows the antimicrobial activity of the invention'scompositions on bacterial, bacteriophage, viral, fungal and nematodemicrobes, using a wide variety of agricultural products (including leafyvegetables, fruiting and flowering vegetables, podded vegetables, bulband stem vegetables, root and tuberous vegetables, Rosaceae familyfruits, Bramble fruits, berries, Asian fruit, North America fruit, rawmeat, raw poultry, raw seafood, fresh seeds sprouts, nuts, eggs, freshherbs, dried herbs, spices, animal hides, feathers) and inanimateobjects (such as those encountered in hospitals, food processing plants,residential buildings, office buildings, etc.).

Furthermore, the data demonstrate that, surprisingly, the invention'scompositions are effective antimicrobials even at their freezingtemperatures, as shown by the reduction of E. coli on shrimp that hasbeen in contact with ice cubes containing the invention's compositions.

Moreover, Table 6 demonstrates the surprising differential effect of theinvention's compositions on pathogenic bacteria on the one hand, and onnon-pathogenic bacteria and bacteriophage on the other hand. Forexample, the data show higher antimicrobial activity against both thepathogenic bacteria E. coli (Gram-negative) and Staphylococcus aureus(Gram-positive) compared to the non-pathogenic Lactobacillus(Gram-positive) and bacteriophage T4.

Table 6 also demonstrates that the invention's compositions are suitableantimicrobials for application to human skin.

EXAMPLE 3 Larger Scale Testing of Antimicrobial Activity

This example was used to demonstrate large batch processing beforecutting or chopping of produce. Testing procedures were done inaccordance to Example 1. Two hundred pounds of each of spinach, springmix and iceberg lettuce were inoculated with E. coli. ATCC Number 43888and the vegetables were tested using immersion and a bubbler tank Theresults are shown in Table 8.

TABLE 8 Antimicrobial activity (log reduction) of a combination of threecomponents E. Coli (ATCC 43888) inoculum 5.51-log Spinach leaf SpringMix^((b)) Iceberg Lettuce (200 pounds) (200 pounds) (200 pounds)Component^((a)) 2 min.^((c)) 30 min.^((c)) 180 min.^((c)) 2 min.^((c))30 min.^((c)) 180 min.^((c)) 2 min.^((c)) 30 min.^((c)) 180 min.^((b))(a1) 4.1 5 5.51 4.4 5.1 5.51 4.3 5.22 5.51 Hypochlorous Acid 0.01 wgt. %& Phosphoric Acid 1 wgt. % & Propylene Glycol 2 wgt. % (a2) 3.5 4.585.51 3.6 5 5.51 3.44 5 5.51 Hydrogen Peroxide 2 wgt. % & Phosphoric Acid1 wgt. % & Propylene Glycol 2 wgt. % ^((a))Components were dissolved in350 liters of potable water and tested at about 33° F. ^((b))Spring mixmay contain any combination of two or more of baby lettuce, greens,endive and radicchio. ^((c))The vegetables were washed in the solutioncontaining the listed components for the period indicated.

The results demonstrate that the invention's compositions “a1” and “a2”successfully resulted in a 100% reduction of a 5.51-log E. coli inoculumafter 180 minutes of contact in a large scale setting without alteringthe color, texture and/or odor of the treated produce.

EXAMPLE 4 Residual Composition After Drying and/or Washing

This Example demonstrates that residual amounts of the invention'scompositions remain on the treated surface, including after rinsing off,to provide continued antimicrobial activity.

TABLE 9 Residual amounts in parts per million (ppm) of componentComponent Spinach leaf Spring Mix Iceberg Lettuce Propylene Glycol 25 1318 Phosphoric Acid 8 9 5 Hydrogen Peroxide 0 0 0 Hypochlorous Acid “CL2”0.001 0.002 0.001

Table 9 shows the results of HPLC analysis of residual amounts ofcomponents on 5 pounds of each of spinach leaf, spring mix (containingbaby lettuces), and iceberg lettuce following treatment with theinvention's compositions in accordance with Table 8 above., Aandabove. Aflow through wash with 1 liter of distilled water for 60 seconds wasused for HPLC residual tests.

EXAMPLE 5 Applying the Compositions Pre-Harvest

Prior to harvesting fruits or vegetables, compositions (A), (B), (C) or(F) of Table 1 may be used as a pre-harvest treatment. Early in themorning or late in the evening a harvester, tractor, truck, ATV, utilityvehicle or by hand could use a pre-mixed composition in a tank used forspraying, fogging, sprinkling or aerosoling. The fruit or vegetablewould be covered by the composition and allowed to dry to form a film.During this drying time the antimicrobial action would continue. Afterthe composition has dried, the edible film left after the water hasevaporated, contains amounts of diol, acid and/or oxidizer that providecontinued antimicrobial activity and preservative functions.

EXAMPLE 6 Applying the Compositions During Harvesting

During harvesting of agricultural of the agricultural products,compositions (A), (B), (C), (D), (E), (F) or (G) of Table 1 may be usedas a harvesting treatment. A harvester, tractor, truck, ATV, utilityvehicle or by hand could use a pre-mixed composition in a tank used forspraying, fogging, sprinkling or aerosoling during picking procedures.If a tow behind or driven harvester is used then a dunking or immersionsystem maybe implemented. Where the tubs, crates, bins, boxes orcontainer with fruits or vegetables may be placed into a tank, basin ortrough. This is done toto thoroughly cover the fruits or vegetables inthe composition for a reasonable amount of time to ensure completecontact. After the harvesting is completed and the load of fruits orvegetables is shipped, the composition dries to form a film. During thisdrying time the antimicrobial activity would continue. After thecomposition has dried, the edible film left after the water hasevaporated, contains amounts of diol, acid and/or oxidizer that providecontinued antimicrobial activity and preservative functions.

EXAMPLE 7 Applying the Compositions Before Washing and Bagging orCrating

When fruits or vegetables are picked or harvested, they are generallytaken to a cold storage facility for chilling before they aretransported for additional processing or directly to food services orstores. Before transporting, the fruits or vegetables are generallypacked into plastic containers that have openings around all four sidesfor air circulation. The containers are easily handled by forklift orother mechanical means. As the forklift removes the containers andplaces them into a storage (such as cold storage) area, the forkliftcould carry a tank with a premixed composition of (A), (B), (C), (D),(E), (F) or (G) of Table 1 and a spray apparatus to spray thecomposition on the fruits or vegetables as the container is being moved.Once the container has been moved to the cooling station facility, aseparate dipping or dousing station could be used to immerse thecontainer into composition (A), (B), (C), (D), (E), (F), (G), (H) or (I)of Table 1 to cover the fruits or vegetables completely. The immersiontank could use a chemical handling system that utilizes a Microprocessor(analog or digital), pH electrode, Orp electrode, Dissolved Oxygenelectrode, Free Chlorine electrode or DPD system. The system would use asignal representing the available composition to turn on chemical pumps,valves or auger to add additional chemicals and or water to makecompositions for use in the tank.

After the container has been immersed it would be removed to drain anddry. During this drying time the antimicrobial activity would continue.After the composition has dried, the edible film left after the waterhas evaporated, contains amounts of diol, acid and/or oxidizer thatprovide continued antimicrobial activity and preservative functions.

EXAMPLE 8 Applying the Compositions During Washing and Bagging orCrating

Once the fruits or vegetables have been brought into the washing andpackaging facility they are typically conveyed either by a belt, flumeor motorized table. During this transit the fruits or vegetables arewashed with either aqueous sprays or aqueous submersion. The composition(A), (B), (C), (D), (E), (F),(G), (H) or (I) of Table 1 may be part ofthe aqueous solution used to clean the fruits or vegetables. Thissolution may also contain various oxidizers, defoamers, surfactants,oils, pesticides, dirt, and or bbugs. The composition could either mixinto the added makeup water or the recycled water. In either case thecomposition could be inserted as combined ingredients or as independentingredients. The composition, either combined or independent, could bepump fed or gravity fed into a suction side of a pump, tank inlet,vessel inlet, trough inlet, flume inlet that is used for recycle ormakeup water, or into the main supply line for the sprays or for thetransport submersion. A microprocessor, analog or digital, pH electrode,Orp electrode, Dissolved Oxygen electrode, Free Chlorine electrode orDPD system may be used for automatic control. The system would use asignal representing the available composition to turn on chemical pumps,valves or auger to add additional chemicals. Alternatively, apre-mixture of the composition could be added based on time, flow,level, turbidity, bio-load, microbial load, etc.

After the fruits or vegetables have been washed they are usually rinsedoff. The composition could be in the rinse water. After the fruits orvegetables are rinsed they are usually dried, then metered to apackaging station. Alternately, prior to the packaging station, thecomposition could be sprayed, fogged or aerosoled, and the compositionis allowed to dry into a very thin film. The edible film left after thewater has evaporated contains amounts of diol, acid and/or oxidizer toprovide continued antimicrobial activity and preservative functions.

EXAMPLE 9 Shelf Life of Oxidizer

This Example was carried out to determine the stability of the exemplaryoxidizer hypochlorous acid in the presence and absence of diol and atdifferent pH. The level of hypochlorous acid was determined using astandard DPD assay.

TABLE 10 Hypochlorous acid stability and shelf life Chlorine (ppm)Minutes Composition “x1a”^((a)) Chlorine Solution^((b)) 1 50 50 15 50 5020 50 30 40 50 45 38 60 35 50 80 33 90 27 50 100 25 115 22 125 20 135 18150 13 170 10 190 7 50 220 3 240 0 400 50 800 50 1600 48 2000 45 2300 432700 41 3000 40 3500 38 4300 36 ^((a))Composition “x1a” is referred toin Table 2, and contains ortho-Phosphoric acid 0.03 wgt. %, propyleneglycol 0.02 wgt. %, hypochlorous acid 0.005 wgt. % at pH 4 (FIG. 1A).^((b))“Chlorine solution” contains ortho-phosphoric acid 0.001 wgt. %and hypochlorous acid 0.005 wgt. % at pH 7 (FIG. 1B).The data in Table 10 and FIG. 1 show that hypochlorous acid in solutionwas less stable in the presence of diol at pH 4, reaching 0% at about 4hours, compared to 100% in the absence of diol at pH 7.

Each and every publication and patent mentioned in the abovespecification is herein incorporated by reference in its entirety forall purposes. Various modifications and variations of the describedmethods and system of the invention will be apparent to those skilled inthe art without departing from the scope and spirit of the invention.Although the invention has been described in connection with specificembodiments, the invention as claimed should not be unduly limited tosuch specific embodiments. Indeed, various modifications of thedescribed modes for carrying out the invention which are obvious tothose skilled in the art and in fields related thereto are intended tobe within the scope of the following claims.

1-22. (canceled)
 23. A method for reducing the number of microbes on asurface of an agricultural product, comprising a) providing i) anagricultural product having a surface comprising a first number ofmicrobes, and ii) an antimicrobially effective amount of a compositionhaving acidic pH and having a temperature from below said composition'sfreezing temperature to 120° C., wherein said composition is selectedfrom the group consisting of 1) a first composition comprising from 0.01to 0.10 wgt. % phosphoric acid and from 0.01 to 1 wgt. % propyleneglycol, and 2) a second composition comprising phosphoric acid,propylene glycol, and chlorine, and b) contacting said surface with saidcomposition under conditions that produce a contacted surface comprisinga reduced number of said microbes compared to said first number of saidmicrobes on said first surface in the absence of said contacting. 24.The method of claim 23, wherein said surface comprises a pathogenicmicrobe and a non-pathogenic microbe, and said reduced number of saidmicrobes comprises a greater reduction in the number of said pathogenicmicrobe than in the number of said non-pathogenic microbe.
 25. Themethod of claim 23, wherein said composition comprises said firstcomposition, and said method further comprises c) drying said contactedsurface to produce a dried surface comprising one or more of saidphosphoric acid and said propylene glycol.
 26. The method of claim 23,wherein said composition comprises said second composition, and saidmethod further comprises c) drying said contacted surface to produce adried surface comprising one or more of said phosphoric acid, of saidpropylene glycol, and of said chlorine.
 27. The method of claim 23,wherein said composition comprises said first composition, and saidmethod further comprises c) washing said contacted surface underconditions that reduce the amount of one or more of said phosphoric acidand of said propylene glycol on said contacted surface.
 28. The methodof claim 23, wherein said composition comprises said second composition,and said method further comprises c) washing said contacted surfaceunder conditions that reduce the amount of one or more of saidphosphoric acid, of said propylene glycol, and of said chlorine on saidcontacted surface.
 29. (canceled)
 30. A method for reducingdiscoloration of an agricultural product, comprising a) providing i) anagricultural product having a surface, ii) an anti-discolorationeffective amount of a composition having acidic pH, said composition isselected from the group consisting of 1) a first composition comprisingfrom 0.01 to 0.10 wgt. % phosphoric acid and from 0.01 to 1 wgt. %propylene glycol, and 2) a second composition comprising phosphoricacid, propylene glycol, and chlorine, and b) contacting saidagricultural product with said composition under conditions that producea contacted surface having reduced discoloration compared todiscoloration of said surface in the absence of said contacting.
 31. Themethod of claim 23, wherein said composition further comprises sodiumlauryl sulfate.
 32. The composition of claim 31, wherein saidcomposition comprises from 0.001 wgt. % to 0.1 wgt. % of said sodiumlauryl sulfate.
 33. The composition of claim 23, wherein said firstcomposition comprises 0.03 wgt. % phosphoric acid, 0.02 wgt. % propyleneglycol, and 0.005 wgt. % chlorine.
 34. The method of claim 30, whereinsaid first surface comprises a first number of pathogenic microbe and asecond number of non-pathogenic microbe, and wherein said contactedsurface comprises i) a reduced number of said pathogenic microbecompared to said first number of pathogenic microbe on said surface inthe absence of said contacting, and ii) a reduced number of saidnon-pathogenic microbe compared to said second number of non-pathogenicmicrobe on said surface in the absence of said contacting.
 35. Themethod of claim 30, wherein said composition comprises said firstcomposition, and said method further comprises c) drying said contactedsurface to produce a dried surface comprising one or more of saidphosphoric acid and said propylene glycol.
 36. The method of claim 30,wherein said composition comprises said second composition, and saidmethod further comprises c) drying said contacted surface to produce adried surface comprising one or more of said phosphoric acid, of saidpropylene glycol, and of said chlorine.
 37. The method of claim 30,wherein said composition comprises said first composition, and saidmethod further comprises c) washing said contacted surface underconditions that reduce the amount of one or more of said phosphoric acidand of said propylene glycol on said contacted surface.
 38. The methodof claim 30, wherein said composition comprises said second composition,and said method further comprises c) washing said contacted surfaceunder conditions that reduce the amount of one or more of saidphosphoric acid, of said propylene glycol, and of said chlorine on saidcontacted surface.
 39. The method of claim 30, wherein said compositionfurther comprises sodium lauryl sulfate.
 40. The method of claim 39,wherein said composition comprises from 0.001 wgt. % to 0.1 wgt. % ofsaid sodium lauryl sulfate.
 41. The method of claim 30, wherein saidfirst composition comprises 0.03 wgt. % phosphoric acid, 0.02 wgt. %propylene glycol, and 0.005 wgt. % chlorine.
 42. The method of claim 34,wherein the reduction in the number of said pathogenic microbe isgreater than the reduction in the number of said non-pathogenic microbe.